RARE DISEASE
Germ cell tumor of testis
Germ cell tumor of testis
Germ cell tumor of testis
Synonyms: Testicular germ cell tumor
Synonyms: Testicular germ cell tumor
Synonyms: Testicular germ cell tumor
Drug discovery
1
drug
With orphan designation
Overview
Germ cell tumors (GCTs) account for over 95% of testicular cancers, primarily affecting males aged 15–44 years. These tumors arise from germ cell neoplasia in situ (GCNIS) and are classified into seminomas (radiation/chemotherapy-sensitive) and nonseminomas (aggressive, mixed histology). Diagnosis relies on imaging, serum markers (AFP, β-hCG), and histopathology. Platinum-based chemotherapy, radiotherapy, and surgery achieve cure rates exceeding 90% even in advanced stages [1][4][16].
Burden
Incidence increasing by 1%–3% annually globally, with Hispanics projected to surpass non-Hispanic whites in the US by 2026 [2][14].
99% 5-year survival for localized disease; 48%–92% for metastatic cases based on risk stratification [4][16].
Long-term morbidity from treatment includes cardiovascular toxicity, secondary malignancies, and infertility [16][19].
Therapies
Localized disease: Radical orchiectomy ± adjuvant chemotherapy (carboplatin for seminoma) or surveillance [4][8][13].
Metastatic disease: BEP regimen (bleomycin, etoposide, cisplatin) for nonseminomas; radiation or carboplatin for seminomas [6][18][13].
Relapsed/refractory: Salvage high-dose chemotherapy with autologous stem cell transplant or targeted therapies (e.g., paclitaxel-gemcitabine combinations) [3][6][13].
Categories: rare neoplastic diseases, rare transplant-related disorders, rare urogenital diseases
Research Papers
1,997 drug discovery papers about Germ cell tumor of testis, with 1 first-in-class and 1 next-in-class emerging drug candidate forecasted to outperform the average preclinical success rate. Recent publications:
1,997 drug discovery papers about Germ cell tumor of testis, with 1 first-in-class and 1 next-in-class emerging drug candidate forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-09-20 | XPO1 Inhibition by Selinexor Induces Nuclear p53 and p21 Accumulation, Cell-Cycle Arrest and Apoptosis in Testicular Germ Cell Tumors
Abstract Background Testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men. Despite high cure rates with cisplatin-based chemotherapy, resistance and long-term toxicities remain clinical challenges in TGCTs. Selinexor, an FDA-approved XPO1 inhibitor, has demonstrated anti-tumor activity in other cancers, but its potential in TGCTs remains unknown. Objective We investigated the antitumor effects of selinexor in TGCT cell lines. Materials and methods XPO1 RNA expression, protein abundance and localization were evaluated by immunohistochemistry in TGCT tissue microarrays and non-neoplastic testicular tissues, and by meta-analysis of publicly available gene-expression microarray datasets from normal testis, primary TGCT specimens as well as TGCT cell lines. Selinexor effects on cell viability, cell-cycle distribution and apoptosis were evaluated using XTT assay and flow cytometry. p53 and p21 expression and localization were analyzed by Western blotting and Immunofluorescence. Results XPO1 expression was heterogeneous across normal testis, TGCT tissues and cell lines. Yet, selinexor reduced TGCT cell viability and induced G1 or G2/M cell-cycle arrest, apoptosis, increased total p53 and p21 protein levels, and their nuclear accumulation. Of note, control fibroblast exhibited limited sensitivity to selinexor. Discussion and conclusion Our findings demonstrate that inhibition of XPO1 with selinexor might be potential therapeutic strategy for TGCTs.
2026-09-09 | G-quadruplex targeting by CX-5461 as a novel therapeutic strategy in testicular germ cell tumors
Abstract Testicular germ cell tumors (TGCTs) are highly curable malignancies; however, resistance to cisplatin-based therapy remains a critical clinical challenge. G-quadruplexes (G4s) are specific guanine rich, four-stranded DNA structures that are enriched at distinct genomic regions. Due to their stability, G4s can influence genome stability, gene expression, DNA replication as well as telomere maintenance. G4 stabilization by small molecules has been shown to have anticancer effects in several tumors but remains underexplored in TGCTs. Here, we systematically evaluated the effect of six different G4- ligands (CX-5461, CX-3543, BRACO19, PDS, CM03 and RHPS4) on TGCT cell lines and non-malignant fibroblasts. Among those G4 ligands, CX-5461 showed strongest cellular changes in TGCTs. We observed dose- and time-dependent G4 stabilization upon CX-5461 treatment in TGCT cells, accompanied by impaired RNA polymerase I-dependent transcription and induction of DNA double-strand breaks, whereas only minimal effects were observed in control fibroblasts. These effects led to the activation of p53 signaling, and consequently an upregulation of canonical p53 target genes, including CDKN1A, MDM2, PIDD1, GADD45A, ZMAT3, SESN1 and PPM1D, resulting in G2/M cell-cycle arrest and apoptosis. The presence of potential quadruplex forming sequences (PQSs) in the promoters or gene bodies of these genes suggests that G4 structures may contribute to the p53 transcriptional response. Collectively, these data demonstrate that CX-5461 appears as highly potent G4-ligand inducing p53-dependent molecular cascades leading to cell death in germ cell tumors. Unlike in many other tumors, p53 remains functionally intact in TGCTs, suggesting that CX-5461-mediated activation of the p53 pathway could be selectively targeted as a therapeutic strategy in TGCTs.
2026-09-03 | In vivo and FACS-based CRISPR/Cas9 screening uncovers VENTX as a crucial element in maintaining seminoma stemness.
Seminoma, which is the most frequent testicular germ cell tumor, has stem cell-like features related to treatment resistance and recurrence, yet the molecular mechanisms that preserve its stemness are not well understood. This study aimed to discover the tumorigenesis and stemness regulation of seminoma. In vivo CRISPR/Cas9 knockout library screening was performed using Tcam-2 seminoma cells implanted into NSG mice to identify tumorigenesis drivers. Parallel FACS-based screening enriched CD117/CD133 double-negative cells to assess the regulators of seminoma stemness. Functional validation included clonogenicity assays, sphere formation tests, and tumor initiation experiments in vivo. RNA sequencing analyzed downstream pathways. In vivo and FACS-based CRISPR/Cas9 screening uncovers VENTX as a crucial element in maintaining seminoma stemness. Functional validation showed that VENTX knockout decreased clonogenicity, sphere formation, and CD117+/CD133+ populations in Tcam-2 cells, and also hindered tumor initiation in vivo. RNA sequencing identified a connection between VENTX and pluripotency pathways, demonstrating a downregulation of ID1/2/3 (essential differentiation inhibitors) and an imbalance in Wnt/β-catenin and TGF-β signaling. VENTX expression showed a significant association with DNA methylation-based stemness scores in testicular germ cell tumors. In conclusion, VENTX is a key regulator of seminoma stemness, presenting a promising target for therapy to address recurrence and resistance in seminoma.
2026-07-10 | Metastatic Testicular Germ Cell Tumor in a Patient With Neurofibromatosis Type 1: Treatment With Trametinib Based on NF1 Gene Mutation.
Neurofibromatosis type 1 (NF1) is a genetic disorder that increases the risk of various tumors. However, its association with testicular germ cell tumors (GCTs) is rare. We report a case of metastatic GCT in an NF1 patient treated with precision medicine. A 40-year-old male with NF1 presented with a left testicular non-seminomatous GCT and lung metastases. The tumor showed progressive disease after chemotherapy (VIP, TGP) and surgical removal of metastases. Comprehensive genomic profiling (CGP) test found a pathogenic NF1 frameshift mutation (p.Q514fs*43). Based on this finding, the patient received the MEK inhibitor trametinib under patient-proposed healthcare services (BELIEVE Trial). Treatment resulted in normalization of serum alpha-fetoprotein (AFP) for 6 months before the tumor eventually progressed. The patient died at 39 months after diagnosis. This is the first report of trametinib for a metastatic GCT with an NF1 mutation, suggesting a possible but limited treatment effect. The prospective trial of patient-proposed healthcare services with multiple targeted agents based on the results of gene profiling by multigene panel test (BELIEVE), NCCH1901/jRCTs031190104.
2026-03-03 | Discovery of natural apigenin analogues as lysine-specific demethylase 1 inhibitors against tumoral testicular germ cells.
Testicular germ cell tumors (TGCT) pose a threat to men's health, discovery of small molecule compounds with potent antiproliferative activity against testicular germ cells is of great significance. Lysine-specific demethylase 1 (LSD1) is overexpressed in tumoral testicular germ cell lines, and it is a promising target for developing agents against TGCT. A series of natural apigenin analogues were evaluated for their inhibitory activity against LSD1, and their structure activity relationships (SARs) were explored. Among them, 8,3'-diprenylapigenin exhibited the most potent inhibitory activity against LSD1 with an IC50 value of 3.60 µM. 7-hydroxy of 8,3'-diprenylapigenin formed a hydrogen bond with Ala809 of LSD1, demonstrating that 7-hydroxy was a dominant group. In addition, 8,3'-diprenylapigenin reversibly and selectively inhibited LSD1 in a concentration-dependent and time-dependent manner. It inhibited proliferation against tumoral testicular germ cell lines NCCIT and NTERA-2 with IC50 values of 9.37 µM and 5.26 µM, respectively. Mechanism studies revealed that 8,3'-diprenylapigenin induced the generation of ROS, inhibited the activity of catalase and decreased the level of ATP in NTERA-2 cells. Meanwhile, it also activated the release of LDH, increased the activity of SOD and enhanced the level of MDA in NTERA-2 cells. These findings indicated that 8,3'-diprenylapigenin is a novel reversible LSD1 inhibitor and deserves further exploration to treat testicular germ cell tumors.
antibodies
2026-08-21 | Pandemic-Era Stage Migration, Registry-Recorded Primary-Site Surgery, and Survival in Early-Onset Testicular Germ Cell Tumors: A SEER Population-Based Study
Background: This study evaluated temporal changes in stage distribution, registry-recorded primary-site surgery, survival outcomes, and exploratory short-term mortality prediction among patients with early-onset testicular germ cell tumors, defined in this study as tumors diagnosed in patients aged 15–49 years, across the study periods. Materials and Methods: This retrospective cohort study used Incidence-SEER Research Data, 17 Registries, November 2025 Submission. Male patients aged 15–49 years diagnosed with malignant testicular germ cell tumors between 2018 and 2023 were included. Diagnosis periods were categorized as the pre-COVID-19 period (2018–2019), COVID-19 period (2020–2021), and 2022–2023 period. Logistic Regression was used to assess factors associated with regional or distant-stage disease. Overall survival was evaluated using Cox regression, while testicular cancer-specific death was additionally evaluated using Aalen–Johansen cumulative incidence estimates and Fine–Gray competing-risk regression. Follow-up was also restricted to 12 months in sensitivity analyses. Machine learning models were developed as an exploratory secondary analysis of 12-month all-cause mortality. Results: A total of 13,303 patients were included. Among patients with known stage, regional or distant-stage disease increased from 31.2% before the pandemic to 33.5% during the COVID-19 period and 34.4% during the 2022–2023 period. In adjusted analysis, COVID-19 (OR = 1.11, 95% CI: 1.02–1.22) and diagnoses during the 2022–2023 period (OR = 1.16, 95% CI: 1.06–1.28) were associated with higher odds of regional or distant-stage disease. In full follow-up analyses, diagnosis during the 2022–2023 period was associated with higher all-cause mortality (HR = 1.29, 95% CI: 1.01–1.65) and testicular cancer-specific death in both cause-specific Cox (HR = 1.49, 95% CI: 1.12–2.00) and Fine–Gray models (sHR = 1.45, 95% CI: 1.09–1.93). In 12-month analyses, the overall survival association was attenuated, whereas the association with testicular cancer-specific death remained significant in the Fine–Gray model (sHR = 1.46, 95% CI: 1.03–2.07). Conclusions: Early-onset testicular germ cell tumors diagnosed during the COVID-19 period and in 2022–2023 showed a modest shift toward regional or distant-stage presentation, while diagnosis in 2022–2023 was associated with higher testicular cancer-specific mortality in competing-risk analyses.
2026-08-09 | Therapeutic targeting of folate receptor alpha (FRα) by antibody-polymer-drug conjugate in testicular germ cell tumors.
Antibody-drug conjugates (ADCs) are an effective treatment for recurrent/refractory testicular germ cell tumors (rrTGCTs). In this exploratory study, we report disease stabilization achieved with the ADC Mirvetuximab soravtansine (MIRV) in a heavily pretreated patient with rrTGCT and compare this ADC with a novel antibody-polymer-drug conjugate (APDC) targeting folate receptor α (FRα), MIRV-P-MMAE. APDC MIRV-P-MMAE with a drug-to-antibody ratio ∼23, was synthesized using orthogonal attachment of a highly hydrophilic copolymer to the hinge region of anti-FRα antibody. Its efficacy was compared with MIRV in vitro in TGCT cells and in vivo in a metastatic model of human testicular choriocarcinoma (CHC). Furthermore, we evaluated FRα expression in TGCT and assessed efficacy of MIRV in an rrTGCT patient. Our investigations revealed the highest antiproliferative effect of MIRV-P-MMAE in FRα-positive CHC cells, JAR and JEG3, with IC50 values of 19.8 pM and 22.5 pM, respectively. These IC50 values are 210- and 70-fold lower than those of MIRV. Cytotoxicity was also confirmed in SuSa, PA1, and NCR-G1 cell lines. High efficacy was demonstrated in animals intravenously injected with JAR-luc cells, in which APDC treatment significantly prolonged survival (p = 0.0002) and completely cured 3 of 8 animals. Our data showed FRα membrane staining in 5,1% of TGCT samples, but confirmed cytoplasmic positivity in all 5 viable rrTGCT post-chemotherapy patient samples, suggesting that APDC with high DAR could be explored in rrTGCT patients with low expression of the target antigen. Our data warrant further exploration of FRα-targeting APDC as a novel approach for treating FRα-expressing rrTGCT.
2026-07-23 | Data from Comprehensive Profiling of Cell Surface Proteins in Testicular Germ Cell Tumors
<div>Abstract<p>To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCT), we retrospectively analyzed archival formalin-fixed, paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0 to 3+) and the percentage of positive tumor cells were used to derive H-scores (0–300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. Claudin-6 (CLDN6) was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. Epidermal growth factor receptor (EGFR) showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3%, ≥100; mean H-score, 212). TROP2 was markedly upregulated in mixed GCTs (78.9%, ≥100; mean, 194.7), particularly in teratoma components (86.7%, ≥100). Nectin-4 and glycoprotein nonmetastatic melanoma B (GPNMB) were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. The enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings, including metastatic, posttreatment, and relapsed or refractory disease.</p>Significance:<p>Relapsed or refractory testicular GCT remains a clinical challenge with limited treatment options. We evaluated cell surface antigens of primary GCTs and identified CLDN6, EGFR, and TROP2 as candidate targets for antigen-directed approaches. These findings provide a rationale for future validation in metastatic and posttreatment disease settings.</p></div>
2026-06-29 | Comprehensive Profiling of Cell Surface Proteins in Testicular Germ Cell Tumors
To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCTs), we retrospectively analyzed archival formalin-fixed paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0-3+) and percentage of positive tumor cells were used to derive H-scores (0-300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. CLDN6 was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. EGFR showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3% ≥100; mean H-score 212.0). TROP2 was markedly upregulated in mixed GCTs (78.9% ≥100; mean 194.7), particularly in teratoma components (86.7% ≥100). Nectin-4 and GPNMB were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. Enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings including metastatic, post-treatment, and relapsed or refractory disease.
2026-05-14 | TNFSF10: a promising prognostic biomarker and therapeutic target for immunotherapy in testicular germ cell tumors.
Testicular germ cell tumors (TGCTs) are malignant neoplasms with a poor prognosis, and the absence of reliable biomarkers for patient stratification and diagnosis presents a significant challenge. We employed an integrated analysis of Single-Cell RNA-Sequencing and TCGA data to evaluate TNFSF10 as a potential biomarker for prognosis and immunotherapy in TGCTs. Our findings revealed that aberrant TNFSF10 expression was significantly associated with patient survival outcomes. Silencing TNFSF10 inhibited the proliferation, migration, and invasion of TGCT cells, highlighting its role in tumor progression. Moreover, TNFSF10 expression was closely correlated with various components of the tumor microenvironment, suggesting its involvement in both the biological behavior of TGCTs and their response to treatment. TNFSF10 emerges as a promising diagnostic and prognostic biomarker for stratification and targeted therapy in TGCTs, offering new prospects for personalized treatment strategies.
cell therapies
2026-03-10 | Salvage Therapy for Relapsed/Refractory Testicular Germ Cell Tumors: King Faisal Hospital and Research Center, Riyadh Experience.
Optimal salvage treatment for relapsed/refractory germ cell tumors in men remains controversial. The options include conventional-dose chemotherapy (CDCT) and high-dose chemotherapy with autologous stem cell transplantation (HDCT-ASCT). There is currently a lack of data from the Middle East on this topic. This study presents the largest single-institution experience in the region. We retrospectively examined patients aged 14 years or older, treated at King Faisal Hospital and Research Center, Riyadh, between 2010 and 2022. Patients previously received first-line, cisplatin-based chemotherapy. The treatment consisted of either CDCT or HDCT-ASCT. The primary endpoints were event-free survival (EFS) and overall survival (OS). Study included 46 patients, of whom 24 and 22 received CDCT and HDCT-ASCT, respectively. Using the IPFSG prognostic model as per local institutional practice, patients in the very low, low, and intermediate risk groups received CDCT. In the 24-patient CDCT group, 16 (66%) were high/very high risk, and only 2 (8%) demonstrated primary refractoriness. Contrarily, of the 22 patients who received HDCT-ASCT, 20 (91%) were classified as high/very high risk, and 17 (77%) were primary refractory to first-line treatment. The HDCT-ASCT cohort was heavily weighted toward high/very high risk. HDCT-ASCT group (59%) had a higher complete remission rate than the CDCT group (59% vs 41%). With a median follow-up of 35 months, the 3-year OS was 60% in the CDCT group and 53% in the HDCT-ASCT group, whereas the 3-year EFS rates were 54% and 43%, respectively. Results indicated that HDCT-ASCT achieved a superior response rate compared with CDCT, despite an adverse prognostic profile and numerically comparable survival outcomes with the lower-risk cohort treated with CDCT. These findings suggest that the selection of higher-risk patients for a more intensive therapy is an effective strategy. However, direct comparison is limited by the retrospective nature.
2025-06-01 | P-001 Fertility among testicular cancer survivors at a male infertility clinic
Abstract Study question Sperm quality is often already abnormal at diagnosis in testicular cancer subjects. We evaluated the fertility of testicular cancer survivors in our male infertility clinic. Summary answer Testicular cancer survivors even with azoospermia can make fatherhood through ICSI following precise evaluation and optimal sperm retrieval surgery. What is known already General improvements in the prognosis of patients with cancer have been achieved through recent advances in surgery, radiotherapy, and chemotherapy treatments, thus increasing the number of patients surviving therapy. However, anti-cancer treatment has detrimental effect on spermatogenesis and testicular cancer inevitably results in there being a single testicle following treatment, subsequent fertility will be a concern. Cryopreservation of ejaculated sperm before cancer treatment is considered to be essential, however it is rarely performed. In contrast, the use of cryopreserved ejaculated sperm for infertility treatment actually occurs in limited number of patients. Study design, size, duration From 2004 through 2023, 3361 male infertility patients consulted our clinic, 117 (3.5%) of whom had a history of cancer treatment. Thirty-nine patients diagnosed with testicular cancer were evaluated for cancer treatment, semen findings, and infertility treatment. Participants/materials, setting, methods The median age was 36 years, the duration from termination of cancer treatment was 5.5 years, and the duration of infertility was 3.0 years. Seminoma was observed in 29 participants, and non-seminomatous germ cell tumor was in 10 participants. Main results and the role of chance Twenty patients were treated with high orchidectomy alone. Whereas three patients were treated with orchidectomy followed by chemotherapy and sixteen participants underwent subsequent retroperitoneal lymph node dissection (RPLND). Semen analyses revealed azoospermia in 26 participants, whereas oligoasthenozoospermia in 13. Although none of the these oligoasthenozoospermic patients achieved spontaneous pregnancy, three achieved pregnancy and delivery through ICSI. Sperm retrieval surgery was attempted in 18 participants with azoospermia. 13 participants underwent microscopic testicular sperm extraction (micro-TESE), two participants received onco-TESE, two patients received microscopic sperm aspiration (MESA), and one patient received retrograde vasal sperm aspiration (ReVSA). Successful motile sperm recovery was observed in 15 participants. 8 deliveries were obtained through ICSI using surgically retrieved sperm. Limitations, reasons for caution Although patients with azoospermia were informed of the need for sperm retrieval surgery, several participants did not choose to undergo the operation. Wider implications of the findings Testicular cancer survivors, even those with azoospermia, can achieve fatherhood through ICSI, after precise evaluation and appropriate sperm retrieval surgery. Trial registration number No
2024-12-27 | TESTICULAR SPARING SURGERY IN SMALL TESTICULAR LESIONS: FUNCTIONAL AND ONCOLOGICAL OUTCOMES
ABSTRACT Objective: To evaluate our testis-sparing surgery ( TSS) experiences in our clinic and outline its oncological and functional outcomes. Materials and Methods: Patients who underwent TSS due to testicular mass in our clinic between 2008 and 2023 were included in the study. Patients with a mass in solitary testis, bilateral testicular mass as well as patients with a normal contralateral testis and a mass of less than 2 cm or less than 30% of the testicular volume were included in the study. Patient demographics, tumor characteristics, and follow-up data were collected and analyzed statistically. Results: A total of 26 patients who underwent TSS were evaluated. Germ Cell Tumor (GCT ) was detected in 9 patients, and benign testicular mass was detected in 17 patients. The mean patient age was 25±6.1 (18-69) years. The mean tumor size was 12.9±4.4 (range 7-24) mm in all patients. Patients with GCTs were followed up for a mean of 21.8±7.8 (10-36) months. Local recurrence was observed in one patient during follow-up, and radical orchiectomy was performed. No recurrence or metastasis was observed in other patients during the follow-up period. Benign lesions were followed up for 21.5±9.3 (10-38) months. Recurrence was not observed. There was no significant decrease in testosterone levels after surgery (p=0.3). Conclusions: Excellent outcomes for benign tumors using TSS were obtained in the present study. TSS could be suggested as a safe and effective treatment option in patients with germ cell tumors, as in the present study. However, more data regarding the potential risks and benefits of TSS with a larger patient series is needed. Keywords: Bilateral testicular tumor, germ cell tumor, solitary testis, testis sparing surgery
2024-07-09 | Follow-Up of Patients Diagnosed With Germinal Testicular Tumors (Seminomas and Non-seminomatous) Treated With a Bone Marrow Transplant and a High Dose of Chemotherapy.
Germinal testicular tumors are the most common malignant neoplasm in men around 20 to 34 years. Even though they are unusual, they have increased incidence in the last decade; they have an excellent prognosis and overall survival at five years, approximately 95%. Divergent data exists regarding treatment options in patients with first, second, and third relapses with conventional therapy. Some studies describe the possible benefit of using high-dose chemotherapy associated with a bone marrow transplant with variable results. The present study describes clinical outcomes, clinical response, mortality, overall survival, and progression-free survival to two years in a group of patients with germinal malignant tumors, seminoma versus non-seminomatous with evidence of progression of the disease at first, second, or third conventional chemotherapy regimens, and who received high dose chemotherapy and bone marrow transplantation at the National Cancer Institute between 2010 and 2021. A retrospective observational study of case series showed that 57% of patients in third-line therapy received high-dose chemotherapy and bone marrow transplantation, with progression disease median time from diagnosis more than two years. Patients in the post-graft period presented infectious complications (71%). The most common were febrile neutropenia (29%) with a mortality rate of 71% (n=5), progression-free survival of 2.3 months, and overall survival of 7.4 months. These results show that in this group of patients, regimens with high-dose chemotherapy associated with bone marrow transplants, have a worse prognosis compared to other cohorts of patients, and may not be the best candidates for this rescue therapy.
2024-05-26 | Ex-vivo microscopic oncotesticular sperm extraction: step-by-step surgical technique at time of radical orchiectomy.
To demonstrate the intraoperative surgical techniques required for simultaneous radical orchiectomy and microscopic oncotesticular sperm extraction (m-OncoTESE) in a step-by-step fashion. Video presentation. University Hospital (University of Chicago). A 37-year-old man (status after right orchiectomy at another institution for stage II-C testicular seminoma with positive preoperative tumor markers) was referred for contralateral orchiectomy of multifocal left testis mass and fertility preservation. Semen analysis before, microscopic testicular sperm extraction during, and semen or testicular specimen analysis after the first orchiectomy were unable to identify any sperm. A postoperative analysis of the m-OncoTESE performed on the left testis resulted in the cryopreservation of 200,000 motile sperm for future assisted reproductive technology (i.e., in vitro fertilization or in vitro fertilization-intracytoplasmic sperm injection). Left radical orchiectomy and left m-OncoTESE. A comprehensive visual documentation of m-OncoTESE surgical techniques with concurrent commentary detailing the reasons behind each surgical step. A brief discussion on the background of m-OncoTESE and alternative fertility preservation methods accompanies the procedure. This video provides a step-by-step guide to performing an m-OncoTESE (proceeding a radical orchiectomy in a patient with testicular cancer) as a means of fertility preservation in an azoospermic patient. Successful extraction and cryopreservation of testicular spermatozoa were achieved after targeted ex-vivo testicular microdissection. Sperm extraction via m-OncoTESE is a viable option for azoospermic patients with testicular cancer undergoing radical orchiectomies. The use of preoperative imaging and microsurgical techniques facilitates and optimizes surgical dissection and sperm recovery.
oligonucleotides
2026-06-29 | MicroRNAs and immunotherapy in testicular germ cell tumors: opportunities and challenges for modulation of the immune microenvironment.
Testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men and, despite high cure rates with cisplatin-based multimodal therapy, a clinically relevant subset of patients develops relapsed, platinum-refractory, or treatment-resistant disease with limited salvage options and substantial long-term morbidity. This unmet need has renewed interest in immunotherapy, yet the experience in TGCT has been notably less successful than in other malignancies. Although TGCTs often display PD-L1 expression, tumor-infiltrating lymphocytes, and other features suggestive of immune engagement, immune checkpoint inhibitors have produced largely disappointing results in unselected patients, underscoring the complexity of immune regulation in this immune-privileged disease. Recent conceptual advances place microRNAs (miRNAs) at the center of this problem. Beyond their established diagnostic and prognostic value, miRNAs are increasingly recognized as upstream regulators of immune checkpoints, antigen-presentation pathways, cytokine signaling, macrophage polarization, dendritic-cell and T-cell function, and extracellular-vesicle-mediated tumor-immune crosstalk. This positions miRNAs not only as biomarkers of disease activity, but also as plausible determinants of immunotherapy sensitivity or resistance. At the same time, the field remains marked by important controversies: PD-L1 expression alone is an unreliable predictor of response; the relative contribution of low tumor mutational burden, testicular immune privilege, and microenvironmental suppression remains unresolved; and many proposed TGCT-associated miRNAs with an immunoregulatory role are still inferential rather than causally validated in disease-specific models. Accordingly, major gaps persist in defining which miRNAs are true functional drivers of immune escape in seminomatous versus non-seminomatous TGCT, how they interact with therapy-induced tumor evolution, and which delivery platforms can achieve safe, tumor-directed modulation. We argue that the next phase of the field should move beyond descriptive biomarker studies toward mechanism-based, TGCT-specific translational strategies integrating miRNA mimics, antagomirs, extracellular vesicles, or miRNA-augmented cellular therapies with immune-priming approaches such as epigenetic therapy, radiotherapy, vaccines, or CAR-T platforms. Such a framework could enable more precise biological stratification and improve the efficacy, durability, and tolerability of immunotherapy in refractory TGCT.
2025-08-30 | Functional Role of miR-138-5p and miR-200b-3p in Testicular Germ Cell Tumors: Molecular Insights into Seminoma and Teratoma Pathogenesis.
This study aims to investigate the molecular mechanisms underlying germ cell tumors (GCTs), focusing specifically on seminomas and teratomas. By analyzing gene expression profiles and miRNA interactions, the goal is to identify key regulatory miRNAs and signaling pathways that differentiate these tumor types and could serve as important regulators for therapy development. Raw data for seminomas and teratomas were extracted from the GEO database, and gene hubs were identified using STRING and Gephi. Signaling pathways and functional annotations were analyzed using miRPathDB, while miRNA-gene interactions were explored via miRWalk. Hub miRNAs were filtered and confirmed using miRDB. This study highlights significant changes in gene expression diversity between tumor and normal gonadal tissues, providing insights into the molecular dynamics of seminomas and teratomas. Distinctions between seminomas and teratomas were identified, shifting the focus toward miRNAs to discover more precise and novel therapeutic approaches. The hub genes of seminomas and teratomas were identified separately. MiRNAs targeting these hub genes were also determined and confirmed. These miRNAs collectively influence essential oncogenic pathways-confirming hsa-miR-138-5p as a regulator of pathways such as Hippo signaling, transcriptional misregulation in cancer, and microRNA cancer signaling in seminomas, and hsa-miR-200b-3p as a regulator of p53 signaling, T cell receptor signaling, and pathways including PI3K/AKT, MAPK/ERK, and Wnt/β-catenin in teratomas-confirming their potential as promising candidates for subtype-specific therapeutic intervention. MiRNAs identified through bioinformatics analyses, and their predicted regulatory roles in key oncogenic pathways, represent potential therapeutic targets or regulators of biological processes. However, further experimental validation is needed to confirm these findings.
2024-12-21 | The New Tumour Biomarker miRNA-371-3p Influences Cisplatin Sensitivity of Testicular Germ Cell Tumour Cell Lines.
Cisplatin is used to treat a variety of malignancies, including testicular germ cell tumours (TGCTs). Although cisplatin-based chemotherapy yields high response rates, a subset of patients develop cisplatin resistance, limiting treatment options and worsening prognosis. Therefore, there is a high clinical need for new therapeutic strategies targeting cisplatin-resistant TGCTs. MicroRNA-371a-3p (miR-371), the new serum biomarker for TGCTs, shows significantly increased expression in cisplatin-resistant TGCT cell lines compared to sensitive parental cell lines. However, the functional impact of miR-371 on cisplatin sensitivity has not been investigated yet. To evaluate the impact of miR-371 on cisplatin sensitivity, antagomirs were used to inhibit miR-371 expression, resulting in a > 98% decrease in miR-371 expression. Cisplatin sensitivity was significantly increased after miR-371 inhibition in cisplatin-resistant and corresponding parental TGCT cell lines, indicating a strongly reduced viability and increased apoptosis after cisplatin treatment in miR-371-inhibited cells. Our results suggest that miR-371 may contribute to the development of cisplatin resistance in TGCTs. Interfering with miR-371 expression can increase the cisplatin sensitivity of tumour cells, which may represent a promising approach to improve future therapeutic outcomes in patients with TGCTs, especially those with cisplatin-resistant disease.
2024-11-13 | Replenishing co-downregulated miR-100-5p and miR-125b-5p in malignant germ cell tumors causes growth inhibition through cell cycle disruption.
MicroRNAs (miRNAs) are short, nonprotein-coding RNAs, and their expression is dysregulated in malignant germ cell tumors (GCTs). Here, we investigated the causes and consequences of downregulated miR-99a-5p/miR-100-5p (functionally identical) and miR-125b-5p levels in malignant GCTs regardless of age, site, or subtype. Quantitative RT-PCR was used to assess miR-99a-5p/miR-100-5p, miR-125b-5p, and associated gene expression in malignant GCT tissues/cell lines [seminoma (Sem), yolk sac tumor (YST), embryonal carcinoma (EC)]. Cells were treated with demethylating 5-azacytidine and pyrosequencing was performed. Combination miR-100-5p/miR-125b-5p mimic replenishment was used to treat malignant GCT cells. Global messenger RNA (mRNA) targets of the replenished miRNAs were identified and Metascape used to study pathway effects. We found that expression levels of miR-99a-5p/miR-100-5p and miR-125b-5p, their respective pri-miRNAs, and associated genes from chromosomes 11 and 21 (chr11/chr21) were downregulated and highly correlated in malignant GCT cells. Treatment with 5-azacytidine caused upregulation of these miRNAs, with pyrosequencing revealing hypermethylation of their chr11/chr21 loci, likely contributing to miR-100-5p/miR-125b-5p downregulation. Combination miR-100-5p/miR-125b-5p mimic replenishment resulted in growth inhibition in Sem/YST cells, with miR-100-5p/miR-125b-5p mRNA targets enriched in downregulated genes, which were involved in cell cycle (confirmed by flow cytometry) and signaling pathways. Knockdown of the miR-100-5p/miR-125b-5p target tripartite motif containing 71 (TRIM71kd) recapitulated miR-100-5p/miR-125b-5p replenishment, with growth inhibition and cell cycle disruption of Sem/YST/EC cells. Further, replenishment led to reduced lin-28 homolog A (LIN28A) levels and concomitant increases in let-7 (MIRLET7B) tumor suppressor miRNAs, creating a sustained reversion of cell phenotype. In summary, combination miR-100-5p/miR-125b-5p mimic replenishment or TRIM71kd caused growth inhibition in malignant GCT cells via cell cycle disruption. Further studies are now warranted, including mimic treatment alongside conventional platinum-based chemotherapy.
2024-11-11 | Single-cell landscape identified SERPINB9 as a key player contributing to stemness and metastasis in non-seminomas
Embryonal carcinoma (EC), characterized by a high degree of stemness similar to that of embryonic stem cells, is the most malignant subtype within non-seminomatous testicular germ cell tumors (TGCTs). However, the mechanisms underlying its malignancy remain unknown. In this study, we employed single-cell RNA sequencing to analyze four non-seminoma samples. Our differential expression analysis revealed high expression of SERPINB9 in metastatic EC cells. We conducted in vitro experiments to further investigate SERPINB9's role in the progression of EC. Functionally, the knockdown of SERPINB9 in NCCIT and NTERA-2 leads to a diminished migratory capability and decreased cis-platin resistance, as demonstrated by Transwell migration assay and drug sensitivity assay. Moreover, embryoid bodies showed reduced size and lower OCT4 expression, alongside heightened expression of differentiation markers AFP, ACTA2, and CD57 in shSERPINB9 cells. In vivo, the role of SERPINB9 in maintaining cancer stemness was validated by the limiting dilution assay. Mechanistically, Bulk RNA-seq further showed downregulation of ERK1/2 signaling and WNT signaling pathways with concomitant upregulation of differentiation pathways subsequent to SERPINB9 knockdown. Additionally, the analysis indicated increased levels of cytokines linked to tertiary lymphoid structures (TLS), such as IL6, IL11, IL15, CCL2, CCL5, and CXCL13 in shSERPINB9 cells, which were further validated by ELISA. Our research indicates that SERPINB9 plays a key role in driving tumor progression by enhancing tumor stemness and suppressing TLS. This study stands as the first to elucidate the molecular signature of non-seminomas at a single-cell level, presenting a wealth of promising targets with substantial potential for informing the development of future therapeutic interventions.
other
2025-11-28 | The expression, regulation, and function of human endogenous retroviruses in genitourinary cancers
Human endogenous retroviruses (HERVs), constituting approximately 8% of the human genome, represent genomic remnants of ancestral retroviral infections that colonized the germline through evolutionary processes. While most HERVs remain epigenetically silenced, their reactivation through environmental stimuli or epigenetic dysregulation enables participation in oncogenesis via viral mimicry, immunomodulation, and insertional mutagenesis. Substantial evidence now implicates aberrant HERVs activity across urologic malignancies-including prostate cancer, renal cell carcinoma (RCC), bladder cancer, and testicular germ cell tumors-where cancer-type-specific mechanisms drive tumor development and progression. These encompass androgen-responsive HERV-K activation in prostate malignancies, hypoxia-inducible factor-mediated ERV immunogenicity in RCC, HERV-derived microRNA silencing of tumor suppressors in bladder cancer, and DNA hypomethylation-associated HERV expression in testicular germ cell tumors. This review synthesizes fundamental HERV biology with recent advances in their diagnostic and therapeutic applications for urologic neoplasms. Key clinical translations include ERV-based stratification models predicting immune checkpoint inhibitor response in metastatic RCC, HERV-E-targeted adoptive T cell therapies, and noncoding RNA biomarkers for early bladder cancer detection. We further discuss unresolved mechanistic paradoxes such as contradictory prognostic associations between HERV superfamily expression and PBRM1 inactivation in RCC, concluding with priorities for future research: validating HERV-derived neoantigens in immunotherapy platforms, optimizing epigenetic priming strategies to enhance viral mimicry effects, and establishing standardized HERV signatures as clinical biomarkers through multi-institutional cohorts.
2025-08-06 | C‐mannosylation promotes ADAMTS1 activation and secretion in human testicular germ cell tumor NEC8 cells
C ‐mannosylation is a protein glycosylation that regulates the functions of target proteins. Although it has been reported that a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1), an important spermatogenesis factor, is C ‐mannosylated, the roles of C ‐mannosylation in ADAMTS1 in testicular cells are still unclear. In this study, we found that ADAMTS1 is C ‐mannosylated at Trp 562 and Trp 565 in testis germ NEC8 cells. To determine the roles of C ‐mannosylation in ADAMTS1, we established cells expressing a C ‐mannosylation‐defective ADAMTS1, in which C ‐mannosylated tryptophan residues were replaced with phenylalanine residues (ADAMTS1/2WF). Processing and secretion of ADAMTS1/2WF were both inhibited compared to those of wild‐type. Moreover, wild‐type ADAMTS1 degraded aggrecan, whereas ADAMTS1/2WF could not. These results indicate the impact of C ‐mannosylation on ADAMTS1 function.
2025-05-28 | Use of CRISPR-KO screening to identify novel genes and pathways potentially contributing to cisplatin resistance in testicular germ cell tumors.
e17020 Background: Testicular germ cell tumors (TGCT) are the most common malignancy in young adult men, with a globally increasing incidence. While these tumors are highly curable due to their exceptional sensitivity to cisplatin (cDDP), a proportion of patients develops a resistance to this chemotherapy, which results in disease progression and patient death. The cause of cisplatin resistance in TGCT has not been elucidated yet and is supposed to be multifactorial. Methods: We conducted a CRISPR Knockout (KO) screening in a TGCT cell line (NCCIT) using a lentiviral sgRNAs library (Addgene #101926) targeting 3015 genes involved in cell cycle regulation and cancer development. The screening population was cultured with or without cDDP (1.25 uM) for 7 days. Samples of treated and control cells were harvested before and after cDDP exposure, the sgRNA cassettes were sequenced by massive parallel sequencing, and the differences in sgRNA abundance were determined using the MAGeCK/MAGeCK-Flute pipeline. Results: The CRISPR-KO screening data analysis identified 59 gene KOs that were significantly positively selected and 7 gene KOs that were significantly negatively selected after cDDP exposure. Among the top positive hits were genes involved in the regulation of apoptosis ( BMF, CDIP1 and BAX ), epigenetic modifiers ( HDAC8, SSX2 ), G-protein regulators ( ADCY8, GNG10 ), and others (e.g. MAGEA2, ACADM, LAMP1 ) — inactivation of these genes potentially contributes to the development of cDDP resistance. Among the top negative hits were genes implicated in DNA damage repair ( FANCB , ERCC5 ), sphingolipid metabolism ( KDSR ), and cytokinesis ( SPECC1L ) — inactivation of these genes may increase sensitivity to cDDP. Conclusions: The results highlight several potential mechanisms of cDPP resistance in TGCT. Some of these genes and pathways are well known and have been previously described in association with platinum resistance in malignant tumors, while others — such as the G-protein regulators, sphingolipid synthesis, and SPECC1L — are novel in this context and may reflect the unique biology of TGCT progression. SPECC1L is involved in spindle organization and microtubule stabilization; its paralog SPECC1 is highly expressed in testis. Polyenoic sphingolipids are essential for the completion of meiosis in male germ cells and spermatozoa production. ADCY8 controls the production of cAMP, which is a key signaling molecule in spermatogenesis. They have also been described in association with malignant proliferation and progression. The identified hits will be validated in subsequent experiments.
2024-06-01 | Decade of insights: Bleomycin-induced pneumonitis and use of G-CSF in chemotherapy-treated germ cell testicular tumors and Hodgkin lymphoma—Real-world outcomes from an academic cancer center.
e17013 Background: Bleomycin induced pulmonary toxicity (BIP) occurs in 4-10% of treated patients. Preclinical reports suggest G-CSF use may increase BIP risk, due to neutrophil migration to lung tissues. Limited real-world clinical data is available and G-CSF use may reduce risk of treatment related toxicity with curative bleomycin combination treatment. Methods: We conducted a retrospective study of patients with GCT (Germ cell tumor) and HL (Hodgkin’s lymphoma) receiving bleomycin containing chemotherapy and at-least on dose of G-CSF at The James Cancer Center from 2010-2022 using the institutional database. BIP was diagnosed mainly using clinical symptoms complemented by radiological evidence when possible. We reviewed 361 patients, 145 in GCT group and 216 in HL group and clinically relevant information was recorded.Descriptive statistics for the study variables were presented using frequencies and percentages overall. The bivariate associations between G-CSF use and clinical outcomes were assessed using chi-square tests or fisher’s exact tests for the overall sample, and by chemotherapy type. We used SPSS 26.0 for all the statistical analyses, and an alpha level of 0.05 (two-sided) was used for all the tests. Results: Data on 361 patients, including 145 GCT and 216 HL patients treated with at least one dose of bleomycin was available. The median age was 36.3 years in the GCT group and 44.2 years in the HL group.144 patients in the GCT and 50 patients in the HL group received at least 1 dose of G-CSF. BIP occurred in 4.8% in the GCT and 7.4% of patients in the HL group. Only 2 out of 16 patients developing BIP in the HL group received G-CSF. All patients with BIP developed symptomatic disease. On bivariate analysis, no association was noted between BIP and G-CSF in either group (p-value =1.00 in GCT and p-value = 0.32 in HL). Febrile neutropenia (FN) was reported in 4 patients in the GCT group and 29 in the HL group. In the HL subgroup, 28 (28/29) patients developing FN did not receive G-CSF (p-value=0.0008). All patients who developed FN required hospital admission. Conclusions: Use of G-CSF in patients receiving bleomycin was not associated with an increased incidence of BIP. Incidence of FN was higher in patients not receiving G-CSF leading to hospital admissions. [Table: see text]
2024-04-11 | Testicular niche repair after gonadotoxic treatments: Current knowledge and future directions.
The testicular niche, which includes the germ cells, somatic cells, and extracellular matrix, plays a crucial role in maintaining the proper functions of the testis. Gonadotoxic treatments, such as chemotherapy and radiation therapy, have significantly improved the survival rates of cancer patients but have also been shown to have adverse effects on the testicular microenvironment. Therefore, repairing the testicular niche after gonadotoxic treatments is essential to restore its function. In recent years, several approaches, such as stem cell transplantation, gene therapy, growth factor therapy, and pharmacological interventions have been proposed as potential therapeutic strategies to repair the testicular niche. This comprehensive review aims to provide an overview of the current understanding of testis damage and repair mechanisms. We will cover a range of topics, including the mechanism of gonadotoxic action, repair mechanisms, and treatment approaches. Overall, this review highlights the importance of repairing the testicular niche after gonadotoxic treatments and identifies potential avenues for future research to improve the outcomes for cancer survivors.
small molecules
2026-09-20 | XPO1 Inhibition by Selinexor Induces Nuclear p53 and p21 Accumulation, Cell-Cycle Arrest and Apoptosis in Testicular Germ Cell Tumors
Abstract Background Testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men. Despite high cure rates with cisplatin-based chemotherapy, resistance and long-term toxicities remain clinical challenges in TGCTs. Selinexor, an FDA-approved XPO1 inhibitor, has demonstrated anti-tumor activity in other cancers, but its potential in TGCTs remains unknown. Objective We investigated the antitumor effects of selinexor in TGCT cell lines. Materials and methods XPO1 RNA expression, protein abundance and localization were evaluated by immunohistochemistry in TGCT tissue microarrays and non-neoplastic testicular tissues, and by meta-analysis of publicly available gene-expression microarray datasets from normal testis, primary TGCT specimens as well as TGCT cell lines. Selinexor effects on cell viability, cell-cycle distribution and apoptosis were evaluated using XTT assay and flow cytometry. p53 and p21 expression and localization were analyzed by Western blotting and Immunofluorescence. Results XPO1 expression was heterogeneous across normal testis, TGCT tissues and cell lines. Yet, selinexor reduced TGCT cell viability and induced G1 or G2/M cell-cycle arrest, apoptosis, increased total p53 and p21 protein levels, and their nuclear accumulation. Of note, control fibroblast exhibited limited sensitivity to selinexor. Discussion and conclusion Our findings demonstrate that inhibition of XPO1 with selinexor might be potential therapeutic strategy for TGCTs.
2026-09-09 | G-quadruplex targeting by CX-5461 as a novel therapeutic strategy in testicular germ cell tumors
Abstract Testicular germ cell tumors (TGCTs) are highly curable malignancies; however, resistance to cisplatin-based therapy remains a critical clinical challenge. G-quadruplexes (G4s) are specific guanine rich, four-stranded DNA structures that are enriched at distinct genomic regions. Due to their stability, G4s can influence genome stability, gene expression, DNA replication as well as telomere maintenance. G4 stabilization by small molecules has been shown to have anticancer effects in several tumors but remains underexplored in TGCTs. Here, we systematically evaluated the effect of six different G4- ligands (CX-5461, CX-3543, BRACO19, PDS, CM03 and RHPS4) on TGCT cell lines and non-malignant fibroblasts. Among those G4 ligands, CX-5461 showed strongest cellular changes in TGCTs. We observed dose- and time-dependent G4 stabilization upon CX-5461 treatment in TGCT cells, accompanied by impaired RNA polymerase I-dependent transcription and induction of DNA double-strand breaks, whereas only minimal effects were observed in control fibroblasts. These effects led to the activation of p53 signaling, and consequently an upregulation of canonical p53 target genes, including CDKN1A, MDM2, PIDD1, GADD45A, ZMAT3, SESN1 and PPM1D, resulting in G2/M cell-cycle arrest and apoptosis. The presence of potential quadruplex forming sequences (PQSs) in the promoters or gene bodies of these genes suggests that G4 structures may contribute to the p53 transcriptional response. Collectively, these data demonstrate that CX-5461 appears as highly potent G4-ligand inducing p53-dependent molecular cascades leading to cell death in germ cell tumors. Unlike in many other tumors, p53 remains functionally intact in TGCTs, suggesting that CX-5461-mediated activation of the p53 pathway could be selectively targeted as a therapeutic strategy in TGCTs.
2026-09-03 | In vivo and FACS-based CRISPR/Cas9 screening uncovers VENTX as a crucial element in maintaining seminoma stemness.
Seminoma, which is the most frequent testicular germ cell tumor, has stem cell-like features related to treatment resistance and recurrence, yet the molecular mechanisms that preserve its stemness are not well understood. This study aimed to discover the tumorigenesis and stemness regulation of seminoma. In vivo CRISPR/Cas9 knockout library screening was performed using Tcam-2 seminoma cells implanted into NSG mice to identify tumorigenesis drivers. Parallel FACS-based screening enriched CD117/CD133 double-negative cells to assess the regulators of seminoma stemness. Functional validation included clonogenicity assays, sphere formation tests, and tumor initiation experiments in vivo. RNA sequencing analyzed downstream pathways. In vivo and FACS-based CRISPR/Cas9 screening uncovers VENTX as a crucial element in maintaining seminoma stemness. Functional validation showed that VENTX knockout decreased clonogenicity, sphere formation, and CD117+/CD133+ populations in Tcam-2 cells, and also hindered tumor initiation in vivo. RNA sequencing identified a connection between VENTX and pluripotency pathways, demonstrating a downregulation of ID1/2/3 (essential differentiation inhibitors) and an imbalance in Wnt/β-catenin and TGF-β signaling. VENTX expression showed a significant association with DNA methylation-based stemness scores in testicular germ cell tumors. In conclusion, VENTX is a key regulator of seminoma stemness, presenting a promising target for therapy to address recurrence and resistance in seminoma.
2026-07-10 | Metastatic Testicular Germ Cell Tumor in a Patient With Neurofibromatosis Type 1: Treatment With Trametinib Based on NF1 Gene Mutation.
Neurofibromatosis type 1 (NF1) is a genetic disorder that increases the risk of various tumors. However, its association with testicular germ cell tumors (GCTs) is rare. We report a case of metastatic GCT in an NF1 patient treated with precision medicine. A 40-year-old male with NF1 presented with a left testicular non-seminomatous GCT and lung metastases. The tumor showed progressive disease after chemotherapy (VIP, TGP) and surgical removal of metastases. Comprehensive genomic profiling (CGP) test found a pathogenic NF1 frameshift mutation (p.Q514fs*43). Based on this finding, the patient received the MEK inhibitor trametinib under patient-proposed healthcare services (BELIEVE Trial). Treatment resulted in normalization of serum alpha-fetoprotein (AFP) for 6 months before the tumor eventually progressed. The patient died at 39 months after diagnosis. This is the first report of trametinib for a metastatic GCT with an NF1 mutation, suggesting a possible but limited treatment effect. The prospective trial of patient-proposed healthcare services with multiple targeted agents based on the results of gene profiling by multigene panel test (BELIEVE), NCCH1901/jRCTs031190104.
2026-03-03 | Discovery of natural apigenin analogues as lysine-specific demethylase 1 inhibitors against tumoral testicular germ cells.
Testicular germ cell tumors (TGCT) pose a threat to men's health, discovery of small molecule compounds with potent antiproliferative activity against testicular germ cells is of great significance. Lysine-specific demethylase 1 (LSD1) is overexpressed in tumoral testicular germ cell lines, and it is a promising target for developing agents against TGCT. A series of natural apigenin analogues were evaluated for their inhibitory activity against LSD1, and their structure activity relationships (SARs) were explored. Among them, 8,3'-diprenylapigenin exhibited the most potent inhibitory activity against LSD1 with an IC50 value of 3.60 µM. 7-hydroxy of 8,3'-diprenylapigenin formed a hydrogen bond with Ala809 of LSD1, demonstrating that 7-hydroxy was a dominant group. In addition, 8,3'-diprenylapigenin reversibly and selectively inhibited LSD1 in a concentration-dependent and time-dependent manner. It inhibited proliferation against tumoral testicular germ cell lines NCCIT and NTERA-2 with IC50 values of 9.37 µM and 5.26 µM, respectively. Mechanism studies revealed that 8,3'-diprenylapigenin induced the generation of ROS, inhibited the activity of catalase and decreased the level of ATP in NTERA-2 cells. Meanwhile, it also activated the release of LDH, increased the activity of SOD and enhanced the level of MDA in NTERA-2 cells. These findings indicated that 8,3'-diprenylapigenin is a novel reversible LSD1 inhibitor and deserves further exploration to treat testicular germ cell tumors.
antibodies
2026-08-21 | Pandemic-Era Stage Migration, Registry-Recorded Primary-Site Surgery, and Survival in Early-Onset Testicular Germ Cell Tumors: A SEER Population-Based Study
Background: This study evaluated temporal changes in stage distribution, registry-recorded primary-site surgery, survival outcomes, and exploratory short-term mortality prediction among patients with early-onset testicular germ cell tumors, defined in this study as tumors diagnosed in patients aged 15–49 years, across the study periods. Materials and Methods: This retrospective cohort study used Incidence-SEER Research Data, 17 Registries, November 2025 Submission. Male patients aged 15–49 years diagnosed with malignant testicular germ cell tumors between 2018 and 2023 were included. Diagnosis periods were categorized as the pre-COVID-19 period (2018–2019), COVID-19 period (2020–2021), and 2022–2023 period. Logistic Regression was used to assess factors associated with regional or distant-stage disease. Overall survival was evaluated using Cox regression, while testicular cancer-specific death was additionally evaluated using Aalen–Johansen cumulative incidence estimates and Fine–Gray competing-risk regression. Follow-up was also restricted to 12 months in sensitivity analyses. Machine learning models were developed as an exploratory secondary analysis of 12-month all-cause mortality. Results: A total of 13,303 patients were included. Among patients with known stage, regional or distant-stage disease increased from 31.2% before the pandemic to 33.5% during the COVID-19 period and 34.4% during the 2022–2023 period. In adjusted analysis, COVID-19 (OR = 1.11, 95% CI: 1.02–1.22) and diagnoses during the 2022–2023 period (OR = 1.16, 95% CI: 1.06–1.28) were associated with higher odds of regional or distant-stage disease. In full follow-up analyses, diagnosis during the 2022–2023 period was associated with higher all-cause mortality (HR = 1.29, 95% CI: 1.01–1.65) and testicular cancer-specific death in both cause-specific Cox (HR = 1.49, 95% CI: 1.12–2.00) and Fine–Gray models (sHR = 1.45, 95% CI: 1.09–1.93). In 12-month analyses, the overall survival association was attenuated, whereas the association with testicular cancer-specific death remained significant in the Fine–Gray model (sHR = 1.46, 95% CI: 1.03–2.07). Conclusions: Early-onset testicular germ cell tumors diagnosed during the COVID-19 period and in 2022–2023 showed a modest shift toward regional or distant-stage presentation, while diagnosis in 2022–2023 was associated with higher testicular cancer-specific mortality in competing-risk analyses.
2026-08-09 | Therapeutic targeting of folate receptor alpha (FRα) by antibody-polymer-drug conjugate in testicular germ cell tumors.
Antibody-drug conjugates (ADCs) are an effective treatment for recurrent/refractory testicular germ cell tumors (rrTGCTs). In this exploratory study, we report disease stabilization achieved with the ADC Mirvetuximab soravtansine (MIRV) in a heavily pretreated patient with rrTGCT and compare this ADC with a novel antibody-polymer-drug conjugate (APDC) targeting folate receptor α (FRα), MIRV-P-MMAE. APDC MIRV-P-MMAE with a drug-to-antibody ratio ∼23, was synthesized using orthogonal attachment of a highly hydrophilic copolymer to the hinge region of anti-FRα antibody. Its efficacy was compared with MIRV in vitro in TGCT cells and in vivo in a metastatic model of human testicular choriocarcinoma (CHC). Furthermore, we evaluated FRα expression in TGCT and assessed efficacy of MIRV in an rrTGCT patient. Our investigations revealed the highest antiproliferative effect of MIRV-P-MMAE in FRα-positive CHC cells, JAR and JEG3, with IC50 values of 19.8 pM and 22.5 pM, respectively. These IC50 values are 210- and 70-fold lower than those of MIRV. Cytotoxicity was also confirmed in SuSa, PA1, and NCR-G1 cell lines. High efficacy was demonstrated in animals intravenously injected with JAR-luc cells, in which APDC treatment significantly prolonged survival (p = 0.0002) and completely cured 3 of 8 animals. Our data showed FRα membrane staining in 5,1% of TGCT samples, but confirmed cytoplasmic positivity in all 5 viable rrTGCT post-chemotherapy patient samples, suggesting that APDC with high DAR could be explored in rrTGCT patients with low expression of the target antigen. Our data warrant further exploration of FRα-targeting APDC as a novel approach for treating FRα-expressing rrTGCT.
2026-07-23 | Data from Comprehensive Profiling of Cell Surface Proteins in Testicular Germ Cell Tumors
<div>Abstract<p>To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCT), we retrospectively analyzed archival formalin-fixed, paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0 to 3+) and the percentage of positive tumor cells were used to derive H-scores (0–300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. Claudin-6 (CLDN6) was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. Epidermal growth factor receptor (EGFR) showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3%, ≥100; mean H-score, 212). TROP2 was markedly upregulated in mixed GCTs (78.9%, ≥100; mean, 194.7), particularly in teratoma components (86.7%, ≥100). Nectin-4 and glycoprotein nonmetastatic melanoma B (GPNMB) were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. The enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings, including metastatic, posttreatment, and relapsed or refractory disease.</p>Significance:<p>Relapsed or refractory testicular GCT remains a clinical challenge with limited treatment options. We evaluated cell surface antigens of primary GCTs and identified CLDN6, EGFR, and TROP2 as candidate targets for antigen-directed approaches. These findings provide a rationale for future validation in metastatic and posttreatment disease settings.</p></div>
2026-06-29 | Comprehensive Profiling of Cell Surface Proteins in Testicular Germ Cell Tumors
To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCTs), we retrospectively analyzed archival formalin-fixed paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0-3+) and percentage of positive tumor cells were used to derive H-scores (0-300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. CLDN6 was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. EGFR showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3% ≥100; mean H-score 212.0). TROP2 was markedly upregulated in mixed GCTs (78.9% ≥100; mean 194.7), particularly in teratoma components (86.7% ≥100). Nectin-4 and GPNMB were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. Enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings including metastatic, post-treatment, and relapsed or refractory disease.
2026-05-14 | TNFSF10: a promising prognostic biomarker and therapeutic target for immunotherapy in testicular germ cell tumors.
Testicular germ cell tumors (TGCTs) are malignant neoplasms with a poor prognosis, and the absence of reliable biomarkers for patient stratification and diagnosis presents a significant challenge. We employed an integrated analysis of Single-Cell RNA-Sequencing and TCGA data to evaluate TNFSF10 as a potential biomarker for prognosis and immunotherapy in TGCTs. Our findings revealed that aberrant TNFSF10 expression was significantly associated with patient survival outcomes. Silencing TNFSF10 inhibited the proliferation, migration, and invasion of TGCT cells, highlighting its role in tumor progression. Moreover, TNFSF10 expression was closely correlated with various components of the tumor microenvironment, suggesting its involvement in both the biological behavior of TGCTs and their response to treatment. TNFSF10 emerges as a promising diagnostic and prognostic biomarker for stratification and targeted therapy in TGCTs, offering new prospects for personalized treatment strategies.
cell therapies
2026-03-10 | Salvage Therapy for Relapsed/Refractory Testicular Germ Cell Tumors: King Faisal Hospital and Research Center, Riyadh Experience.
Optimal salvage treatment for relapsed/refractory germ cell tumors in men remains controversial. The options include conventional-dose chemotherapy (CDCT) and high-dose chemotherapy with autologous stem cell transplantation (HDCT-ASCT). There is currently a lack of data from the Middle East on this topic. This study presents the largest single-institution experience in the region. We retrospectively examined patients aged 14 years or older, treated at King Faisal Hospital and Research Center, Riyadh, between 2010 and 2022. Patients previously received first-line, cisplatin-based chemotherapy. The treatment consisted of either CDCT or HDCT-ASCT. The primary endpoints were event-free survival (EFS) and overall survival (OS). Study included 46 patients, of whom 24 and 22 received CDCT and HDCT-ASCT, respectively. Using the IPFSG prognostic model as per local institutional practice, patients in the very low, low, and intermediate risk groups received CDCT. In the 24-patient CDCT group, 16 (66%) were high/very high risk, and only 2 (8%) demonstrated primary refractoriness. Contrarily, of the 22 patients who received HDCT-ASCT, 20 (91%) were classified as high/very high risk, and 17 (77%) were primary refractory to first-line treatment. The HDCT-ASCT cohort was heavily weighted toward high/very high risk. HDCT-ASCT group (59%) had a higher complete remission rate than the CDCT group (59% vs 41%). With a median follow-up of 35 months, the 3-year OS was 60% in the CDCT group and 53% in the HDCT-ASCT group, whereas the 3-year EFS rates were 54% and 43%, respectively. Results indicated that HDCT-ASCT achieved a superior response rate compared with CDCT, despite an adverse prognostic profile and numerically comparable survival outcomes with the lower-risk cohort treated with CDCT. These findings suggest that the selection of higher-risk patients for a more intensive therapy is an effective strategy. However, direct comparison is limited by the retrospective nature.
2025-06-01 | P-001 Fertility among testicular cancer survivors at a male infertility clinic
Abstract Study question Sperm quality is often already abnormal at diagnosis in testicular cancer subjects. We evaluated the fertility of testicular cancer survivors in our male infertility clinic. Summary answer Testicular cancer survivors even with azoospermia can make fatherhood through ICSI following precise evaluation and optimal sperm retrieval surgery. What is known already General improvements in the prognosis of patients with cancer have been achieved through recent advances in surgery, radiotherapy, and chemotherapy treatments, thus increasing the number of patients surviving therapy. However, anti-cancer treatment has detrimental effect on spermatogenesis and testicular cancer inevitably results in there being a single testicle following treatment, subsequent fertility will be a concern. Cryopreservation of ejaculated sperm before cancer treatment is considered to be essential, however it is rarely performed. In contrast, the use of cryopreserved ejaculated sperm for infertility treatment actually occurs in limited number of patients. Study design, size, duration From 2004 through 2023, 3361 male infertility patients consulted our clinic, 117 (3.5%) of whom had a history of cancer treatment. Thirty-nine patients diagnosed with testicular cancer were evaluated for cancer treatment, semen findings, and infertility treatment. Participants/materials, setting, methods The median age was 36 years, the duration from termination of cancer treatment was 5.5 years, and the duration of infertility was 3.0 years. Seminoma was observed in 29 participants, and non-seminomatous germ cell tumor was in 10 participants. Main results and the role of chance Twenty patients were treated with high orchidectomy alone. Whereas three patients were treated with orchidectomy followed by chemotherapy and sixteen participants underwent subsequent retroperitoneal lymph node dissection (RPLND). Semen analyses revealed azoospermia in 26 participants, whereas oligoasthenozoospermia in 13. Although none of the these oligoasthenozoospermic patients achieved spontaneous pregnancy, three achieved pregnancy and delivery through ICSI. Sperm retrieval surgery was attempted in 18 participants with azoospermia. 13 participants underwent microscopic testicular sperm extraction (micro-TESE), two participants received onco-TESE, two patients received microscopic sperm aspiration (MESA), and one patient received retrograde vasal sperm aspiration (ReVSA). Successful motile sperm recovery was observed in 15 participants. 8 deliveries were obtained through ICSI using surgically retrieved sperm. Limitations, reasons for caution Although patients with azoospermia were informed of the need for sperm retrieval surgery, several participants did not choose to undergo the operation. Wider implications of the findings Testicular cancer survivors, even those with azoospermia, can achieve fatherhood through ICSI, after precise evaluation and appropriate sperm retrieval surgery. Trial registration number No
2024-12-27 | TESTICULAR SPARING SURGERY IN SMALL TESTICULAR LESIONS: FUNCTIONAL AND ONCOLOGICAL OUTCOMES
ABSTRACT Objective: To evaluate our testis-sparing surgery ( TSS) experiences in our clinic and outline its oncological and functional outcomes. Materials and Methods: Patients who underwent TSS due to testicular mass in our clinic between 2008 and 2023 were included in the study. Patients with a mass in solitary testis, bilateral testicular mass as well as patients with a normal contralateral testis and a mass of less than 2 cm or less than 30% of the testicular volume were included in the study. Patient demographics, tumor characteristics, and follow-up data were collected and analyzed statistically. Results: A total of 26 patients who underwent TSS were evaluated. Germ Cell Tumor (GCT ) was detected in 9 patients, and benign testicular mass was detected in 17 patients. The mean patient age was 25±6.1 (18-69) years. The mean tumor size was 12.9±4.4 (range 7-24) mm in all patients. Patients with GCTs were followed up for a mean of 21.8±7.8 (10-36) months. Local recurrence was observed in one patient during follow-up, and radical orchiectomy was performed. No recurrence or metastasis was observed in other patients during the follow-up period. Benign lesions were followed up for 21.5±9.3 (10-38) months. Recurrence was not observed. There was no significant decrease in testosterone levels after surgery (p=0.3). Conclusions: Excellent outcomes for benign tumors using TSS were obtained in the present study. TSS could be suggested as a safe and effective treatment option in patients with germ cell tumors, as in the present study. However, more data regarding the potential risks and benefits of TSS with a larger patient series is needed. Keywords: Bilateral testicular tumor, germ cell tumor, solitary testis, testis sparing surgery
2024-07-09 | Follow-Up of Patients Diagnosed With Germinal Testicular Tumors (Seminomas and Non-seminomatous) Treated With a Bone Marrow Transplant and a High Dose of Chemotherapy.
Germinal testicular tumors are the most common malignant neoplasm in men around 20 to 34 years. Even though they are unusual, they have increased incidence in the last decade; they have an excellent prognosis and overall survival at five years, approximately 95%. Divergent data exists regarding treatment options in patients with first, second, and third relapses with conventional therapy. Some studies describe the possible benefit of using high-dose chemotherapy associated with a bone marrow transplant with variable results. The present study describes clinical outcomes, clinical response, mortality, overall survival, and progression-free survival to two years in a group of patients with germinal malignant tumors, seminoma versus non-seminomatous with evidence of progression of the disease at first, second, or third conventional chemotherapy regimens, and who received high dose chemotherapy and bone marrow transplantation at the National Cancer Institute between 2010 and 2021. A retrospective observational study of case series showed that 57% of patients in third-line therapy received high-dose chemotherapy and bone marrow transplantation, with progression disease median time from diagnosis more than two years. Patients in the post-graft period presented infectious complications (71%). The most common were febrile neutropenia (29%) with a mortality rate of 71% (n=5), progression-free survival of 2.3 months, and overall survival of 7.4 months. These results show that in this group of patients, regimens with high-dose chemotherapy associated with bone marrow transplants, have a worse prognosis compared to other cohorts of patients, and may not be the best candidates for this rescue therapy.
2024-05-26 | Ex-vivo microscopic oncotesticular sperm extraction: step-by-step surgical technique at time of radical orchiectomy.
To demonstrate the intraoperative surgical techniques required for simultaneous radical orchiectomy and microscopic oncotesticular sperm extraction (m-OncoTESE) in a step-by-step fashion. Video presentation. University Hospital (University of Chicago). A 37-year-old man (status after right orchiectomy at another institution for stage II-C testicular seminoma with positive preoperative tumor markers) was referred for contralateral orchiectomy of multifocal left testis mass and fertility preservation. Semen analysis before, microscopic testicular sperm extraction during, and semen or testicular specimen analysis after the first orchiectomy were unable to identify any sperm. A postoperative analysis of the m-OncoTESE performed on the left testis resulted in the cryopreservation of 200,000 motile sperm for future assisted reproductive technology (i.e., in vitro fertilization or in vitro fertilization-intracytoplasmic sperm injection). Left radical orchiectomy and left m-OncoTESE. A comprehensive visual documentation of m-OncoTESE surgical techniques with concurrent commentary detailing the reasons behind each surgical step. A brief discussion on the background of m-OncoTESE and alternative fertility preservation methods accompanies the procedure. This video provides a step-by-step guide to performing an m-OncoTESE (proceeding a radical orchiectomy in a patient with testicular cancer) as a means of fertility preservation in an azoospermic patient. Successful extraction and cryopreservation of testicular spermatozoa were achieved after targeted ex-vivo testicular microdissection. Sperm extraction via m-OncoTESE is a viable option for azoospermic patients with testicular cancer undergoing radical orchiectomies. The use of preoperative imaging and microsurgical techniques facilitates and optimizes surgical dissection and sperm recovery.
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2026-06-29 | MicroRNAs and immunotherapy in testicular germ cell tumors: opportunities and challenges for modulation of the immune microenvironment.
Testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men and, despite high cure rates with cisplatin-based multimodal therapy, a clinically relevant subset of patients develops relapsed, platinum-refractory, or treatment-resistant disease with limited salvage options and substantial long-term morbidity. This unmet need has renewed interest in immunotherapy, yet the experience in TGCT has been notably less successful than in other malignancies. Although TGCTs often display PD-L1 expression, tumor-infiltrating lymphocytes, and other features suggestive of immune engagement, immune checkpoint inhibitors have produced largely disappointing results in unselected patients, underscoring the complexity of immune regulation in this immune-privileged disease. Recent conceptual advances place microRNAs (miRNAs) at the center of this problem. Beyond their established diagnostic and prognostic value, miRNAs are increasingly recognized as upstream regulators of immune checkpoints, antigen-presentation pathways, cytokine signaling, macrophage polarization, dendritic-cell and T-cell function, and extracellular-vesicle-mediated tumor-immune crosstalk. This positions miRNAs not only as biomarkers of disease activity, but also as plausible determinants of immunotherapy sensitivity or resistance. At the same time, the field remains marked by important controversies: PD-L1 expression alone is an unreliable predictor of response; the relative contribution of low tumor mutational burden, testicular immune privilege, and microenvironmental suppression remains unresolved; and many proposed TGCT-associated miRNAs with an immunoregulatory role are still inferential rather than causally validated in disease-specific models. Accordingly, major gaps persist in defining which miRNAs are true functional drivers of immune escape in seminomatous versus non-seminomatous TGCT, how they interact with therapy-induced tumor evolution, and which delivery platforms can achieve safe, tumor-directed modulation. We argue that the next phase of the field should move beyond descriptive biomarker studies toward mechanism-based, TGCT-specific translational strategies integrating miRNA mimics, antagomirs, extracellular vesicles, or miRNA-augmented cellular therapies with immune-priming approaches such as epigenetic therapy, radiotherapy, vaccines, or CAR-T platforms. Such a framework could enable more precise biological stratification and improve the efficacy, durability, and tolerability of immunotherapy in refractory TGCT.
2025-08-30 | Functional Role of miR-138-5p and miR-200b-3p in Testicular Germ Cell Tumors: Molecular Insights into Seminoma and Teratoma Pathogenesis.
This study aims to investigate the molecular mechanisms underlying germ cell tumors (GCTs), focusing specifically on seminomas and teratomas. By analyzing gene expression profiles and miRNA interactions, the goal is to identify key regulatory miRNAs and signaling pathways that differentiate these tumor types and could serve as important regulators for therapy development. Raw data for seminomas and teratomas were extracted from the GEO database, and gene hubs were identified using STRING and Gephi. Signaling pathways and functional annotations were analyzed using miRPathDB, while miRNA-gene interactions were explored via miRWalk. Hub miRNAs were filtered and confirmed using miRDB. This study highlights significant changes in gene expression diversity between tumor and normal gonadal tissues, providing insights into the molecular dynamics of seminomas and teratomas. Distinctions between seminomas and teratomas were identified, shifting the focus toward miRNAs to discover more precise and novel therapeutic approaches. The hub genes of seminomas and teratomas were identified separately. MiRNAs targeting these hub genes were also determined and confirmed. These miRNAs collectively influence essential oncogenic pathways-confirming hsa-miR-138-5p as a regulator of pathways such as Hippo signaling, transcriptional misregulation in cancer, and microRNA cancer signaling in seminomas, and hsa-miR-200b-3p as a regulator of p53 signaling, T cell receptor signaling, and pathways including PI3K/AKT, MAPK/ERK, and Wnt/β-catenin in teratomas-confirming their potential as promising candidates for subtype-specific therapeutic intervention. MiRNAs identified through bioinformatics analyses, and their predicted regulatory roles in key oncogenic pathways, represent potential therapeutic targets or regulators of biological processes. However, further experimental validation is needed to confirm these findings.
2024-12-21 | The New Tumour Biomarker miRNA-371-3p Influences Cisplatin Sensitivity of Testicular Germ Cell Tumour Cell Lines.
Cisplatin is used to treat a variety of malignancies, including testicular germ cell tumours (TGCTs). Although cisplatin-based chemotherapy yields high response rates, a subset of patients develop cisplatin resistance, limiting treatment options and worsening prognosis. Therefore, there is a high clinical need for new therapeutic strategies targeting cisplatin-resistant TGCTs. MicroRNA-371a-3p (miR-371), the new serum biomarker for TGCTs, shows significantly increased expression in cisplatin-resistant TGCT cell lines compared to sensitive parental cell lines. However, the functional impact of miR-371 on cisplatin sensitivity has not been investigated yet. To evaluate the impact of miR-371 on cisplatin sensitivity, antagomirs were used to inhibit miR-371 expression, resulting in a > 98% decrease in miR-371 expression. Cisplatin sensitivity was significantly increased after miR-371 inhibition in cisplatin-resistant and corresponding parental TGCT cell lines, indicating a strongly reduced viability and increased apoptosis after cisplatin treatment in miR-371-inhibited cells. Our results suggest that miR-371 may contribute to the development of cisplatin resistance in TGCTs. Interfering with miR-371 expression can increase the cisplatin sensitivity of tumour cells, which may represent a promising approach to improve future therapeutic outcomes in patients with TGCTs, especially those with cisplatin-resistant disease.
2024-11-13 | Replenishing co-downregulated miR-100-5p and miR-125b-5p in malignant germ cell tumors causes growth inhibition through cell cycle disruption.
MicroRNAs (miRNAs) are short, nonprotein-coding RNAs, and their expression is dysregulated in malignant germ cell tumors (GCTs). Here, we investigated the causes and consequences of downregulated miR-99a-5p/miR-100-5p (functionally identical) and miR-125b-5p levels in malignant GCTs regardless of age, site, or subtype. Quantitative RT-PCR was used to assess miR-99a-5p/miR-100-5p, miR-125b-5p, and associated gene expression in malignant GCT tissues/cell lines [seminoma (Sem), yolk sac tumor (YST), embryonal carcinoma (EC)]. Cells were treated with demethylating 5-azacytidine and pyrosequencing was performed. Combination miR-100-5p/miR-125b-5p mimic replenishment was used to treat malignant GCT cells. Global messenger RNA (mRNA) targets of the replenished miRNAs were identified and Metascape used to study pathway effects. We found that expression levels of miR-99a-5p/miR-100-5p and miR-125b-5p, their respective pri-miRNAs, and associated genes from chromosomes 11 and 21 (chr11/chr21) were downregulated and highly correlated in malignant GCT cells. Treatment with 5-azacytidine caused upregulation of these miRNAs, with pyrosequencing revealing hypermethylation of their chr11/chr21 loci, likely contributing to miR-100-5p/miR-125b-5p downregulation. Combination miR-100-5p/miR-125b-5p mimic replenishment resulted in growth inhibition in Sem/YST cells, with miR-100-5p/miR-125b-5p mRNA targets enriched in downregulated genes, which were involved in cell cycle (confirmed by flow cytometry) and signaling pathways. Knockdown of the miR-100-5p/miR-125b-5p target tripartite motif containing 71 (TRIM71kd) recapitulated miR-100-5p/miR-125b-5p replenishment, with growth inhibition and cell cycle disruption of Sem/YST/EC cells. Further, replenishment led to reduced lin-28 homolog A (LIN28A) levels and concomitant increases in let-7 (MIRLET7B) tumor suppressor miRNAs, creating a sustained reversion of cell phenotype. In summary, combination miR-100-5p/miR-125b-5p mimic replenishment or TRIM71kd caused growth inhibition in malignant GCT cells via cell cycle disruption. Further studies are now warranted, including mimic treatment alongside conventional platinum-based chemotherapy.
2024-11-11 | Single-cell landscape identified SERPINB9 as a key player contributing to stemness and metastasis in non-seminomas
Embryonal carcinoma (EC), characterized by a high degree of stemness similar to that of embryonic stem cells, is the most malignant subtype within non-seminomatous testicular germ cell tumors (TGCTs). However, the mechanisms underlying its malignancy remain unknown. In this study, we employed single-cell RNA sequencing to analyze four non-seminoma samples. Our differential expression analysis revealed high expression of SERPINB9 in metastatic EC cells. We conducted in vitro experiments to further investigate SERPINB9's role in the progression of EC. Functionally, the knockdown of SERPINB9 in NCCIT and NTERA-2 leads to a diminished migratory capability and decreased cis-platin resistance, as demonstrated by Transwell migration assay and drug sensitivity assay. Moreover, embryoid bodies showed reduced size and lower OCT4 expression, alongside heightened expression of differentiation markers AFP, ACTA2, and CD57 in shSERPINB9 cells. In vivo, the role of SERPINB9 in maintaining cancer stemness was validated by the limiting dilution assay. Mechanistically, Bulk RNA-seq further showed downregulation of ERK1/2 signaling and WNT signaling pathways with concomitant upregulation of differentiation pathways subsequent to SERPINB9 knockdown. Additionally, the analysis indicated increased levels of cytokines linked to tertiary lymphoid structures (TLS), such as IL6, IL11, IL15, CCL2, CCL5, and CXCL13 in shSERPINB9 cells, which were further validated by ELISA. Our research indicates that SERPINB9 plays a key role in driving tumor progression by enhancing tumor stemness and suppressing TLS. This study stands as the first to elucidate the molecular signature of non-seminomas at a single-cell level, presenting a wealth of promising targets with substantial potential for informing the development of future therapeutic interventions.
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2025-11-28 | The expression, regulation, and function of human endogenous retroviruses in genitourinary cancers
Human endogenous retroviruses (HERVs), constituting approximately 8% of the human genome, represent genomic remnants of ancestral retroviral infections that colonized the germline through evolutionary processes. While most HERVs remain epigenetically silenced, their reactivation through environmental stimuli or epigenetic dysregulation enables participation in oncogenesis via viral mimicry, immunomodulation, and insertional mutagenesis. Substantial evidence now implicates aberrant HERVs activity across urologic malignancies-including prostate cancer, renal cell carcinoma (RCC), bladder cancer, and testicular germ cell tumors-where cancer-type-specific mechanisms drive tumor development and progression. These encompass androgen-responsive HERV-K activation in prostate malignancies, hypoxia-inducible factor-mediated ERV immunogenicity in RCC, HERV-derived microRNA silencing of tumor suppressors in bladder cancer, and DNA hypomethylation-associated HERV expression in testicular germ cell tumors. This review synthesizes fundamental HERV biology with recent advances in their diagnostic and therapeutic applications for urologic neoplasms. Key clinical translations include ERV-based stratification models predicting immune checkpoint inhibitor response in metastatic RCC, HERV-E-targeted adoptive T cell therapies, and noncoding RNA biomarkers for early bladder cancer detection. We further discuss unresolved mechanistic paradoxes such as contradictory prognostic associations between HERV superfamily expression and PBRM1 inactivation in RCC, concluding with priorities for future research: validating HERV-derived neoantigens in immunotherapy platforms, optimizing epigenetic priming strategies to enhance viral mimicry effects, and establishing standardized HERV signatures as clinical biomarkers through multi-institutional cohorts.
2025-08-06 | C‐mannosylation promotes ADAMTS1 activation and secretion in human testicular germ cell tumor NEC8 cells
C ‐mannosylation is a protein glycosylation that regulates the functions of target proteins. Although it has been reported that a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1), an important spermatogenesis factor, is C ‐mannosylated, the roles of C ‐mannosylation in ADAMTS1 in testicular cells are still unclear. In this study, we found that ADAMTS1 is C ‐mannosylated at Trp 562 and Trp 565 in testis germ NEC8 cells. To determine the roles of C ‐mannosylation in ADAMTS1, we established cells expressing a C ‐mannosylation‐defective ADAMTS1, in which C ‐mannosylated tryptophan residues were replaced with phenylalanine residues (ADAMTS1/2WF). Processing and secretion of ADAMTS1/2WF were both inhibited compared to those of wild‐type. Moreover, wild‐type ADAMTS1 degraded aggrecan, whereas ADAMTS1/2WF could not. These results indicate the impact of C ‐mannosylation on ADAMTS1 function.
2025-05-28 | Use of CRISPR-KO screening to identify novel genes and pathways potentially contributing to cisplatin resistance in testicular germ cell tumors.
e17020 Background: Testicular germ cell tumors (TGCT) are the most common malignancy in young adult men, with a globally increasing incidence. While these tumors are highly curable due to their exceptional sensitivity to cisplatin (cDDP), a proportion of patients develops a resistance to this chemotherapy, which results in disease progression and patient death. The cause of cisplatin resistance in TGCT has not been elucidated yet and is supposed to be multifactorial. Methods: We conducted a CRISPR Knockout (KO) screening in a TGCT cell line (NCCIT) using a lentiviral sgRNAs library (Addgene #101926) targeting 3015 genes involved in cell cycle regulation and cancer development. The screening population was cultured with or without cDDP (1.25 uM) for 7 days. Samples of treated and control cells were harvested before and after cDDP exposure, the sgRNA cassettes were sequenced by massive parallel sequencing, and the differences in sgRNA abundance were determined using the MAGeCK/MAGeCK-Flute pipeline. Results: The CRISPR-KO screening data analysis identified 59 gene KOs that were significantly positively selected and 7 gene KOs that were significantly negatively selected after cDDP exposure. Among the top positive hits were genes involved in the regulation of apoptosis ( BMF, CDIP1 and BAX ), epigenetic modifiers ( HDAC8, SSX2 ), G-protein regulators ( ADCY8, GNG10 ), and others (e.g. MAGEA2, ACADM, LAMP1 ) — inactivation of these genes potentially contributes to the development of cDDP resistance. Among the top negative hits were genes implicated in DNA damage repair ( FANCB , ERCC5 ), sphingolipid metabolism ( KDSR ), and cytokinesis ( SPECC1L ) — inactivation of these genes may increase sensitivity to cDDP. Conclusions: The results highlight several potential mechanisms of cDPP resistance in TGCT. Some of these genes and pathways are well known and have been previously described in association with platinum resistance in malignant tumors, while others — such as the G-protein regulators, sphingolipid synthesis, and SPECC1L — are novel in this context and may reflect the unique biology of TGCT progression. SPECC1L is involved in spindle organization and microtubule stabilization; its paralog SPECC1 is highly expressed in testis. Polyenoic sphingolipids are essential for the completion of meiosis in male germ cells and spermatozoa production. ADCY8 controls the production of cAMP, which is a key signaling molecule in spermatogenesis. They have also been described in association with malignant proliferation and progression. The identified hits will be validated in subsequent experiments.
2024-06-01 | Decade of insights: Bleomycin-induced pneumonitis and use of G-CSF in chemotherapy-treated germ cell testicular tumors and Hodgkin lymphoma—Real-world outcomes from an academic cancer center.
e17013 Background: Bleomycin induced pulmonary toxicity (BIP) occurs in 4-10% of treated patients. Preclinical reports suggest G-CSF use may increase BIP risk, due to neutrophil migration to lung tissues. Limited real-world clinical data is available and G-CSF use may reduce risk of treatment related toxicity with curative bleomycin combination treatment. Methods: We conducted a retrospective study of patients with GCT (Germ cell tumor) and HL (Hodgkin’s lymphoma) receiving bleomycin containing chemotherapy and at-least on dose of G-CSF at The James Cancer Center from 2010-2022 using the institutional database. BIP was diagnosed mainly using clinical symptoms complemented by radiological evidence when possible. We reviewed 361 patients, 145 in GCT group and 216 in HL group and clinically relevant information was recorded.Descriptive statistics for the study variables were presented using frequencies and percentages overall. The bivariate associations between G-CSF use and clinical outcomes were assessed using chi-square tests or fisher’s exact tests for the overall sample, and by chemotherapy type. We used SPSS 26.0 for all the statistical analyses, and an alpha level of 0.05 (two-sided) was used for all the tests. Results: Data on 361 patients, including 145 GCT and 216 HL patients treated with at least one dose of bleomycin was available. The median age was 36.3 years in the GCT group and 44.2 years in the HL group.144 patients in the GCT and 50 patients in the HL group received at least 1 dose of G-CSF. BIP occurred in 4.8% in the GCT and 7.4% of patients in the HL group. Only 2 out of 16 patients developing BIP in the HL group received G-CSF. All patients with BIP developed symptomatic disease. On bivariate analysis, no association was noted between BIP and G-CSF in either group (p-value =1.00 in GCT and p-value = 0.32 in HL). Febrile neutropenia (FN) was reported in 4 patients in the GCT group and 29 in the HL group. In the HL subgroup, 28 (28/29) patients developing FN did not receive G-CSF (p-value=0.0008). All patients who developed FN required hospital admission. Conclusions: Use of G-CSF in patients receiving bleomycin was not associated with an increased incidence of BIP. Incidence of FN was higher in patients not receiving G-CSF leading to hospital admissions. [Table: see text]
2024-04-11 | Testicular niche repair after gonadotoxic treatments: Current knowledge and future directions.
The testicular niche, which includes the germ cells, somatic cells, and extracellular matrix, plays a crucial role in maintaining the proper functions of the testis. Gonadotoxic treatments, such as chemotherapy and radiation therapy, have significantly improved the survival rates of cancer patients but have also been shown to have adverse effects on the testicular microenvironment. Therefore, repairing the testicular niche after gonadotoxic treatments is essential to restore its function. In recent years, several approaches, such as stem cell transplantation, gene therapy, growth factor therapy, and pharmacological interventions have been proposed as potential therapeutic strategies to repair the testicular niche. This comprehensive review aims to provide an overview of the current understanding of testis damage and repair mechanisms. We will cover a range of topics, including the mechanism of gonadotoxic action, repair mechanisms, and treatment approaches. Overall, this review highlights the importance of repairing the testicular niche after gonadotoxic treatments and identifies potential avenues for future research to improve the outcomes for cancer survivors.
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Drug Discovery Landscape
1 orphan drug designation for Germ cell tumor of testis, including 1 approved therapy.
1 orphan drug designation for Germ cell tumor of testis, including 1 approved therapy.
Drug | Therapy type | Orphan designation | Clinical trial status | Sponsor |
|---|---|---|---|---|
Ifosfamide [Ifex] | small molecules | FDA | Approved | Bristol-Myers Squibb Pharmaceutical Research Institute |
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