small molecules

2026-06-19 | Data from β-Catenin–Cohesin Ring–CEGRs/ALCDs Axis Activation Contributes to the Development of Hepatoblastoma and Fibrolamellar HCC

<div>Abstract<p>The pediatric and adolescent liver cancers, hepatoblastoma (HBL) and fibrolamellar hepatocellular carcinoma (FLC), respectively, are dangerous diseases requiring aggressive surgery, when feasible, and nontargeted toxic chemotherapy for a chance of cure due to insufficient knowledge of underlying molecular mechanisms. We have previously reported the essential role of ph-S675-β-catenin in the reorganization of genomic structure in HBL and FLC by oncogenic activation via chromosomal regions called cancer-enhancing genomic regions or aggressive liver cancer domains (CEGR/ALCD). In FLC, the fusion DNAJB1-PKAc (J-PKAc) oncoprotein phosphorylates β-catenin at Ser675, triggering such CEGRs/ALCDs-mediated activation of oncogenes. In this study, we found that all members of the cohesin ring—CTCF, Rad21, SMC1, SMC3, and STAG1—and β-catenin–TCF4 are bound to CEGRs/ALCDs of oncogenes in HBL and FLC, as well as many other cancers, and that this binding increases transcription. Examination of a large cohort of HBL and FLC samples revealed that cohesin ring expression is dramatically elevated in the majority. The cohesin ring, as well as the ph-S675-β-catenin–TCF4–p300 complex, is detected on both the promoter and intron-located CEGRs/ALCDs of <i>NRF2</i> and <i>Thy1</i>, correlating with increased transcription. This suggests that the cohesin ring creates the DNA loop for oncogene activation. The inhibition of the cohesin ring by JQ1 reduces the proliferation of HBL and FLC cells in culture, as well as cells expressing the FLC-specific J-PKAc fusion oncogene.</p>Implications:<p>These studies provide evidence that J-PKAc–β-catenin and the cohesin ring cooperate in oncogenic activation for both HBL and FLC.</p></div>

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2026-05-12 | Metastatic Fibrolamellar Hepatocellular Carcinoma in a Young Adult: A Case Report and Narrative Review.

Fibrolamellar hepatocellular carcinoma (FLC) is a rare and distinct histologic subtype of hepatocellular carcinoma that typically arises in non-cirrhotic livers of adolescents and young adults without underlying viral hepatitis or chronic liver disease. Accounting for less than 1% of all primary liver cancers, FLC is characterized by unique clinical, radiologic, and molecular features, most notably the highly characteristic DNAJB1-PRKACA fusion. Due to its rarity and frequently nonspecific presentation, diagnosis is often delayed or misinterpreted as by benign hepatic lesions such as focal nodular hyperplasia or adenoma. This report presents a rare case of metastatic FLC in a young adult who initially presented with right upper quadrant pain, anemia, and preserved hepatic function. Imaging revealed a large hepatic mass with a central non-enhancing area suggestive of necrosis, and histopathologic evaluation aided in the diagnosis based on characteristic lamellar fibrosis, polygonal eosinophilic cells, and positive CK7 and HepPar-1 staining. Confirmatory molecular testing through DNAJB1-PRKACA fusion was not performed. Planned treatment with a combination regimen of nivolumab and neratinib was scheduled; however, the patient deteriorated rapidly, and systemic therapy could not be initiated before death. Through this case and accompanying review, we aim to highlight the diagnostic complexity, molecular underpinnings, and current therapeutic approaches of FLC. This case was notable for advanced peritoneal carcinomatosis at presentation, severe thrombocytosis, and the diagnostic and therapeutic challenges posed by metastatic fibrolamellar carcinoma without molecular confirmation. Taken together, this underscores the importance of early recognition, molecular testing, and coordinated clinical management in optimizing outcomes for this uncommon malignancy.

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2026-04-03 | Abstract 1002: Parallel drug development strategies for fibrolamellar hepatocellular carcinoma and proteasome-dependent multiple myeloma

Abstract Fibrolamellar Hepatocellular carcinoma (FLC) is a rare liver cancer which occurs primarily in young adults and children under the age of 40. In FLC, a deletion of 400,000 base pairs occurs in one chromosome 19 copy, leading to the formation of a fusion protein comprised of a domain of the heat shock protein DNAJB1 and the catalytic subunit of Protein Kinase A (DNAJB1-PRKACA). Patients are typically asymptomatic at early stages, and 5-year survival remains below 50%. With surgery as the only partially effective treatment and no approved systemic therapies, there is a critical need for new therapeutics. The primary model compound of this project, Napabucasin, is a naphthoquinone natural product that has shown wide spectrum anti-cancer activities. Napabucasin was found to be a top hit in drug screening showing potency against FLC patient derived xenografts (PDX) models and direct-from-patient FLC tumor cells. Napabucasin is known to work in multiple signaling pathways including protein translation, the generation of reactive oxygen species, suppression of tumor cell stemness, and blocking tumor cell apoptotic escape. We anticipate the organic synthesis and development of optimized Napabucasin analogues, along with the evaluation of their cytotoxicity and proliferation effectiveness compared to the parent compound, Napabucasin, will result in compounds with improved bioavailability and efficacy, particularly in FLC model cell lines (Huh7-Chimera). In an additional approach to cancer drug development, we are evaluating the efficacy of promising next generation proteasome inhibitors. In eukaryotic cells, protein homeostasis is maintained through controlled synthesis and degradation, with the Ubiquitin Proteasome System (UPS) serving as the major pathway for regulated proteolysis. Cancer cells increase UPS activity to manage proteotoxic stress from rapid proliferation, leading to degradation of tumor-suppressive proteins and activation of oncogenic pathways. Targeted inhibition of UPS function causes accumulation of ubiquitinated proteins, G2/M arrest, and apoptosis, establishing the proteasome as a validated anticancer target. The 20S core of proteasome contains three catalytic β subunits (β1, β2, β5). Among all three, the chymotrypsin-like β5 activity remains the primary driver of cytotoxicity for clinically approved inhibitors such as bortezomib, carfilzomib; however, their therapeutic use is limited by toxicity. Peptidic β-lactones (the cystargolide family), are promising next-generation proteasome inhibitors. This project evaluates newly synthesized cystargolide analogs for cytotoxicity, bioavailability, and proteasome inhibition in RPMI-8226 myeloma cells. Using cell-based assays and IC50 determination, we aim to establish structure activity relationships and advance optimized β-lactone inhibitors with improved therapeutic potential. Citation Format: Shalini Rawat, Najmeh Rahimi, Jared Alderman, Celeste Villasenor, Rodolfo Tello-Aburto, Barbara Lyons. Parallel drug development strategies for fibrolamellar hepatocellular carcinoma and proteasome-dependent multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 1002.

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2026-04-03 | Utility of Yttrium-90 Radioembolization for the Treatment of Fibrolamellar Hepatocellular Carcinoma: A Tertiary Cancer Center Experience.

To describe the safety and efficacy of Yttrium-90 transarterial radioembolization (Y90-TARE) in patients with fibrolamellar hepatocellular carcinoma (FL-HCC). This is a retrospective study of patients with FL-HCC treated with Y90-TARE in a single tertiary cancer center. Y90-TARE was performed using Y90 glass or resin microspheres. Treatment response was evaluated at 1, 3, and 6 months using the modified Response Evaluation Criteria in Solid Tumors (mRECIST) guidelines. Electronic medical records were retrospectively reviewed to evaluate clinical outcomes, complications, tumor response on imaging, overall survival (OS), and progression-free survival (PFS). MIM Software (v7.1, Cleveland, OH) was used for dosimetry analysis and contouring. Seven patients (5 females, 2 males; median age at the time of Y90-TARE: 24, range: 16-77 years) with FL-HCC underwent nine Y90-TARE procedures. The median progression-free survival (PFS) was 9.0 months, and the median overall survival (OS) was 15.8 months. Partial response (PR) was observed in 86% of patients at 3 months and 100% of evaluable patients at 6 months. One patient underwent successful liver transplantation, and another received hepatectomy after treatment. No adverse events grade 3 or greater were reported within 30 days. One patient developed a hepatic abscess two months post-procedure, and another developed a tumor to duodenum fistula three months after Y90-TARE. The median absorbed tumor dose was 405 Gy (mean: 579 ± 442 Gy; range: 109-1362 Gy), and the median absorbed dose to perfused normal liver was 83 Gy (mean: 96 ± 41 Gy; range: 63.5-190 Gy). The median administered activity was 3.85 GBq, with a median lung shunt fraction of 4.59% and a median lung dose of 6.4 Gy. The study is limited by its retrospective design and small sample size in this rare tumor cohort. Treatment of FL-HCC patients with unresectable tumors with 90Y-TARE is safe and shows a favorable response to treatment.

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2026-03-23 | Fibrolamellar hepatocellular carcinoma: Case report of a rare tumor and a brief review of literature

Fibrolamellar hepatocellular carcinoma (FL-HCC) is a rare primary liver malignancy that typically arises in young adults without underlying cirrhosis or other established hepatic risk factors. We present the case of a 24-year-old male with a four-month history of progressive right upper quadrant abdominal pain, unintentional weight loss, and early satiety. Laboratory evaluation demonstrated mildly elevated liver transaminases and a markedly elevated serum vitamin B12 level of 3,240 pg/mL. Alpha-fetoprotein (AFP) levels were within normal limits. Cross-sectional imaging revealed a 12 cm hypervascular mass within the left hepatic lobe, with a central fibrotic scar and areas of calcification. Core needle biopsy followed by molecular analysis confirmed the diagnosis of fibrolamellar carcinoma through identification of the pathognomonic DNAJB1–PRKACA gene fusion. The patient underwent an extended left hepatectomy (R0 resection) and achieved a disease-free interval of 20 months. Twenty-six months after the initial diagnosis, routine surveillance imaging detected metastatic recurrence involving the lungs and L3 vertebral body. The patient was managed with a multidisciplinary approach that included pulmonary metastasectomy, platinum-based chemotherapy, and participation in a clinical trial evaluating a PRKACA inhibitor. This case underscores the importance of vigilant long-term surveillance in patients with FL-HCC and highlights the emerging role of targeted molecular therapies in addressing recurrent diseases.

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cell therapies

2024-11-25 | Models of fibrolamellar carcinomas, tools for evaluation of a new era of treatments.

Fibrolamellar carcinoma (FLC) is a rare but fatal cancer that occurs primarily in young people. There are currently no known effective treatments, although several promising treatments appear to be in development. Genetic studies have confirmed that almost all FLC tumors have a fusion protein marker (DNAJB1-PRKACA) encoded by a fusion gene (DNAJB1-PRKACA); It is currently accepted as a diagnostic criterion for FLCs. Several research teams have established patient-derived xenograft (PDX) FLC models using immunocompromised animals as hosts and patient tissue samples (tumors or ascites) as primary sources for PDX-derived organoids. These FLC organoids are composed of FLC epithelia, endothelial progenitor cells, and stellate cells. CRISPR/Cas9 was used as a gene editing technique to modify mature hepatocytes to obtain ex vivo FLC-like cells expressing the fusion gene and/or other mutated genes associated with FLCs. Although these models simulate some but not all FLC features. Drug screening using these models has not proven effective in identifying clinically useful treatments. Genetic studies comparing FLCs to normal maturing endodermal cell lineages have shown that FLCs share genetic signatures not with hepatocytes, but with subpopulations of biliary tree stem cells (BTSCs), hepato/pancreatic stem/progenitor cells that consistently reside in peribiliary glands (PBGs) located in the biliary tree and are sources of stem cells for the formation and postnatal regeneration of the liver and pancreas. Therefore, it is expected that models of BTSCs, instead of hepatocytes may prove more useful. In this review, we summarize the status of the various FLC models and their features, applications, and limitations. They provide opportunities to understand the cause and characteristics of this deadly disease and are models from which effective treatments can be identified.

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2024-03-22 | DNAJB1-PRKACA fusion neoantigens elicit rare endogenous T cell responses that potentiate cell therapy for fibrolamellar carcinoma.

Fibrolamellar carcinoma (FLC) is a liver tumor with a high mortality burden and few treatment options. A promising therapeutic vulnerability in FLC is its driver mutation, a conserved DNAJB1-PRKACA gene fusion that could be an ideal target neoantigen for immunotherapy. In this study, we aim to define endogenous CD8 T cell responses to this fusion in FLC patients and evaluate fusion-specific T cell receptors (TCRs) for use in cellular immunotherapies. We observe that fusion-specific CD8 T cells are rare and that FLC patient TCR repertoires lack large clusters of related TCR sequences characteristic of potent antigen-specific responses, potentially explaining why endogenous immune responses are insufficient to clear FLC tumors. Nevertheless, we define two functional fusion-specific TCRs, one of which has strong anti-tumor activity in vivo. Together, our results provide insights into the fragmented nature of neoantigen-specific repertoires in humans and indicate routes for clinical development of successful immunotherapies for FLC.

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2024-01-03 | Treating rare tumors with liver transplantation.

The success of liver transplantation (LT) in treating unresectable hepatocellular carcinoma (HCC) has resulted in interest in LT for other oncologic conditions. Here, we discuss the role of LT for rare oncologic indications including metastatic gastroenteropancreatic neuroendocrine tumors (GEP-NETs), hepatic epitheliod hemangioendothelioma (HEHE), fibrolamellar hepatocellular carcinoma (FLC), and hepatic angiosarcoma (HAS). Conditions reviewed have been documented indications for LT in the available literature. We summarize the experience of LT for these indications and proposed management guidelines. GEP-NETs with isolated metastases to the liver can be treated with LT with excellent long-term outcomes (10-year survival 88%) if strict selection criteria are used (low-intermediate grade, Ki-67% < 20%, complete resection of primary tumor, stable disease for 6 months, <50% hepatic involvement). HEHE is a rare hepatic tumor for which LT can be performed with reported 10-year survival around 70%. FLC is a distinct clinical entity to HCC and is optimally treated with surgical resection though experience with LT is described in observational series (5-year survival 50%, recurrence in 10%). HAS is a rapidly progressive tumor with a dismal prognosis with or without treatment, including LT.

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2023-12-07 | Development of pre-clinical murine models for fibrolamellar hepatocellular carcinoma

ABSTRACT Fibrolamellar hepatocellular carcinoma (FLC) is a rare form of cancer that affects primarily adolescents and young adults. FLC tumors are typically associated with an intrachromosomal deletion resulting in expression of a fusion protein between the chaperone DNAJ1B and the protein kinase PKA. FLC is challenging to study because of its rarity and limited pre-clinical models. Here we developed a novel transgenic mouse model of FLC. In this model, DNAJ1B-PKA expression in the liver of mouse embryos results in perinatal lethality associated with liver developmental defects, while DNAJ1B-PKA expression in the liver of adult mice initiates tumors resembling FLC at low penetrance. We sought to develop ex vivo cell models from these tumors but failed to establish long-term cell lines. New pre-clinical models of FLC will provide novel insights into the biology of this rare cancer and may help identify novel therapeutic strategies.

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2022-11-01 | Surgical therapy and survival in young patients with stage I–II hepatocellular carcinoma: a retrospective cohort study

Hepatocellular carcinoma (HCC) is regarded as a high-mortality cancer, but the effectiveness of surgical strategies for young patients with early-stage HCC remains controversial. We aimed to analyze the survival in young patients with stage I-II HCC who underwent different kinds of surgical treatments.Overall survival (OS) and cancer-specific survival (CSS) were compared among patients aged 18-45 years with stage I-II HCC from the Surveillance, Epidemiology, and End Results (SEER) database (2004-2013) who underwent local tumor destruction (LTD), wedge or segmental resection (WSR), lobectomy resection (LR), liver transplantation (LT), or non-surgery. Univariate and multivariate analyses and Kaplan-Meier method were used to examine the OS and CSS of the patients. A stratification analysis of CSS was also conducted among the subgroups.Data from 664 patients were extracted. The median survival time was 46 months. In the multivariate analysis of OS, compared with non-surgery, LTD [hazard ratio (HR), 0.37; 95% confidence interval (CI): 0.25-0.54; P<0.0001], LR (HR, 0.29; 95% CI: 0.19-0.45; P<0.0001), and WSR (HR, 0.26; 95% CI: 0.17-0.39; P<0.0001) had better outcomes, and LT had the best survival benefit (HR, 0.24; 95% CI: 0.16-0.36; P<0.0001), which was similar to CSS. In the stratification analysis, compared with the non-surgery group, among patients with chemotherapy, LT reduced the risk of CSS by 64% (HR, 0.36; 95% CI: 0.19-0.66; P interaction=0.0004).Surgery offers a survival benefit compared with non-surgery for young patients with stage I-II HCC. LT is associated with better survival than WSR, LR, and LTD.

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antibodies

2026-04-03 | Abstract 2456: Profiling the expression of antibody-drug conjugates in fibrolamellar hepatocellular carcinoma

Abstract Background: Fibrolamellar carcinoma (FLC) is a rare and aggressive primary liver cancer that primarily affects adolescents and young adults. It is biologically distinct from other forms of primary liver cancer, such as hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA). There are no standard or approved systemic therapies for advanced FLC, and most patients present with unresectable disease or experience recurrence after surgery. These factors underscore the need for new therapeutic approaches. Antibody-drug conjugates (ADCs) have shown efficacy in several solid tumors by delivering cytotoxic payloads to tumor-associated surface antigens with improved specificity, but their utility in FLC is not known. To evaluate the relevance of ADC-based therapies in FLC, we assessed the expression of five clinically actionable ADC targets (NECTIN4, TROP2, CLDN18.2, HER2, and B7-H3) in an expanded cohort of FLC specimens. Methods: Archival FFPE samples were obtained from consented patients through the Johns Hopkins Liver Cancer Tissue Bank and the Fibrolamellar Cancer Foundation, including FLC tumors (n=58; 38 primary and 20 metastatic lesions). Immunohistochemistry (IHC) was performed for all five markers. Expression patterns of interest in FLC were compared with control cohorts of HCC (n=10) and CCA (n=10). Three pathologists, blinded to clinical data, independently scored staining using both a 0 to 3 plus ordinal IHC scale and a semi-quantitative H-score. Results: Among the five ADC targets evaluated, only B7-H3 and HER2 showed detectable staining in FLC. NECTIN4, TROP2, and CLDN18.2 were uniformly negative, scored as 0+ in all tumors, and had H-scores of 0. B7-H3 demonstrated the strongest expression. 3+ staining was observed in 20 of 58 tumors (34.5%). Tumor-cell H-scores ranged from 10 to 240, with a mean of 132. B7-H3 was also expressed in the tumor microenvironment, with prominent staining in cancer-associated stroma and fibroblasts. Stromal H-scores ranged from 40 to 300, with a mean of 199, often exceeding tumor-cell expression. HER2 showed detectable 3+ staining in 12 of 58 tumors (21.7%), with H-scores ranging from 2 to 270 and a mean of 70. One patient in this cohort with 3+ HER2 expression received fam-trastuzumab deruxtecan-nxki, and experienced a durable partial response lasting approximately one year. Conclusions: B7-H3 and HER2 are promising ADC targets in FLC. While a subset of patients may already be eligible for fam-trastuzumab deruxtecan-nxki under its tumor-agnostic indication, B7-H3 shows particularly strong and widespread expression across both tumor and stromal compartments. These findings support further preclinical and clinical evaluation of B7-H3- and HER2-directed ADCs as targeted treatment strategies for advanced FLC. Citation Format: Waqar Arif, Elsa Hallab, Franshisca Hayek, Howard Liu Li, Mari Nakazawa, Mark E. Furth, Andrew S. Liss, Patricia Cogswell, Ezra G. Baraban, Jacqueline Birkness-Gartman, Marina Baretti, Robert A. Anders, Mark Yarchoan. Profiling the expression of antibody-drug conjugates in fibrolamellar hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2456.

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2026-02-18 | Procalcitonin as a potential tumor marker for fibrolamellar hepatocellular carcinoma: Insights from three patients and a literature review.

Fibrolamellar hepatocellular carcinoma (fHCC) was diagnosed through magnetic resonance imaging in three young patients (with biopsies confirming the diagnosis in two of them prior to referral) aged 17, 22, and 24 years. All had normal or near-normal standard biochemistry results, except for increased levels of procalcitonin (PCT) and C-reactive protein (CRP) in the absence of any infectious source: PCT: 41, 5.8, and 1.2 ng/mL, respectively (normal range < 0.05 ng/mL) and CRP: 127, 21, and 14 mg/L, respectively (normal range: 0-5 mg/L). Right hepatectomy with negative surgical margins resulted in the eventual normalization of CRP levels in all patients, and PCT levels normalized in the latter two patients. The PCT level was 0.07 ng/mL at 3 months in the first patient. The emergence of two bone metastases in the rib and right femur at 7 months was accompanied by an increase in PCT levels to 0.8 ng/mL. Radiotherapy provided transient control in the femur only. Consequently, a thoracic wall resection was performed, followed by a segmental resection of the femur. Treatment with bevacizumab at a dose of 15 mg/kg/day and atezolizumab at 1200 mg/day every 21 days was started. A PET-CT scan conducted at the 42nd month showed no signs of recurrence, and the PCT level was 0.04 ng/mL. The other patients exhibited normal PCT levels and no evidence of recurrence at 16- and 6-months post-treatment. Since PCT measurement is routinely available, baseline levels should be measured at least once during the workup of patients with fHCC.

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2025-11-03 | Fibrolamellar hepatocellular carcinoma: Advances, challenges and opportunities in a rare malignancy.

Fibrolamellar hepatocellular carcinoma is a rare and unique subtype of primary liver cancer that predominantly affects adolescents and young adults who do not have underlying liver disease or cirrhosis. Representing less than five percent of all liver tumors, it poses significant diagnostic and therapeutic challenges due to its uncommon occurrence, vague clinical symptoms, and absence of standardized treatment protocols. This review summarizes the current understanding of the disease, covering its epidemiology, clinical presentation, imaging characteristics, histopathologic features, molecular biology, and treatment approaches. Patients often present with nonspecific abdominal symptoms, and typical tumor markers, such as alpha-fetoprotein, are usually absent. Imaging may show a central scar and calcifications, while histology reveals large eosinophilic tumor cells separated by fibrous bands. A defining molecular hallmark is the DNAJB1-PRKACA fusion gene, found in the majority of cases. Surgical resection remains the primary curative treatment for localized disease, although recurrence rates are high. Liver transplantation may be an option for selected patients with non-resectable, liver-confined tumors. Treatment options for advanced disease are limited, with some benefit observed from chemotherapy and targeted agents. Recent developments in molecular therapies and immunotherapy offer promise, but further research and clinical trial participation are essential to improve outcomes in this challenging malignancy.

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2025-11-01 | 968 Novel strategies for modulating dendritic cell and T cell responses with bispecific antibodies bound to designed protein nanoparticle cages

Background CD8 + T cells are the prime mediators of effective anti-tumor immunity, which is dependent on recognition of tumor antigens by CD8+ T cells and interactions among CD8+ T cells, dendritic cells (DCs) and CD4+ T cells1; however, CD8+ T cell exhaustion limits proliferation and cytokine production. Reversing CD8+ T cell dysfunction is important for improving endogenous anti-tumor responses, and new methods are needed. Recent studies demonstrated intratumoral clusters of DCs, CD4+ T cells, and CD8+ T cells can lead to more PD1+ CXCL13+ T helper and TCF7+ progenitor CD8+ T cells after immune checkpoint blockade, suggesting interactions between these cells can overcome T cell dysfunction.2 Here, we describe a novel bispecific antibody concept for increasing DC and CD4+ T cell interaction toward reinvigorating anti-tumor responses.Methods Designed protein nanoparticle cages (hAbC) were generated to bind to two Fc portions of antibodies. Two hAbCs were created with antibodies targeting CD3 and CD11c and CD4 and CD11c. hAbCs were cultured with human splenic DCs and CD4 + T cells. Proximity of cells was assessed by live confocal microscopy and flow cytometry. DCs and CD4+ T cells maturation and activation was measured by flow cytometry.Results CD3-CD11c hAbC and CD4-CD11c hAbC were successfully generated and contained one copy of each antibody per antibody cage. DCs and CD4 + T cells co-cultured with CD3-CD11c hAbCs co-localized more than DCs and CD4+ T cells co-cultured without hAbC starting at 30min of culture (150 versus 20), and the colocalization remained stable for 120 mins of culture. At peak, approximately 20% of DCs were within 10 pixels of a CD4+ T cells versus 7% in the control group (figure 1). The number of co-localized DC and CD4+ T cells decreased by 50% at 2h and to baseline by 10h. Similar findings were seen with the CD4-CD11c hAbC. By flow cytometry, doublets of DC and CD4+ T cells were also identified at various doses of hAbC (figure 2). Co-culture of DCs and CD4+ T cells lead to modulation of activation markers on CD4+ T cells and maturation molecules on DCs.Conclusions We describe a novel application of bispecific antibody technology using designed protein nanoparticle cages, hAbC. Co-culture of DCs and CD4 + T cells with the hAbCs led to sustained co-localized of cells and modulation of activation and maturation markers. These results suggest hAbC could be used to augment anti-tumor immunity.References Binnewies M, Roberts EW, Kersten K, et al. Understanding the tumor immune microenvironment (TIME) for effective therapy. Nat Med. 2018;24(5):541–50.Magen A, Hamon P, Fiaschi N, et al. Intratumoral dendritic cell-CD4(+) T helper cell niches enable CD8(+) T cell differentiation following PD-1 blockade in hepatocellular carcinoma. Nat Med. 2023;29(6):1389–99.Ethics Approval This study was approved the Institutional Review Boards (IRB) of Fred Hutch Cancer Center (FHCC IRB 1765) and University of Washington (UW IRB 1852).Abstract 968 Figure 1DC and CD4+ T cells co-localized after culture with hAbC. The number of dendritic cells and T cells within 10 pixels of each other after culture with hAbC or controlAbstract 968 Figure 2DC and CD4+ T cell dose response. hAbC dose response curve for co-localized cells at 18 hours of culture by flow cytometry

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2025-08-03 | Clinicopathological features and treatment outcomes of patients with fibrolamellar hepatocellular carcinoma: a retrospective multicenter study.

Fibrolamellar Hepatocellular Carcinoma (FLHCC) is a rare primary liver tumour that is distinct from conventional HCC in both histopathological and clinical features. There is no clear consensus on its treatment. Assess the demographics, prognosis, and treatment- particularly systemic therapies-of FLHCC patients across multiple centers in Türkiye. Retrospective. Multicenter. Patients with histopathological diagnosis of pure fibrolamellar hepatocellular carcinoma. Treatment responses of metastatic stage patients were evaluated. Progression-free survival (PFS) and overall survival (OS) analyses of metastatic stage first-line treatments were performed. 39 patients with FLHCC. The 5-year survival for all patients was 48%, 80% in stage 1, 57% in stage 2, 53% in stage 3 and 0% in stage 4. The median PFS for chemotherapy (n=10) and sorafenib (n=6) patients in the metastatic stage first series was 5.7 months and 2.8 months, respectively (P=.031). Median OS was 12.1 months for chemotherapy and 8.8 months for sorafenib (P=.853) in the metastatic stage. Conventional chemotherapies, especially gemcitabine and oxaliplatin combination can be used as systemic treatment options. Immunotherapies, chemoimmunotherapy and immunotherapy plus anti vascular endothelial growth factor combinations may be considered by clinicians. Small sample size and the variability of the treatment modalities administered in the patients.

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oligonucleotides

2024-03-21 | DNAJB1-PRKACA fusion protein-regulated LINC00473 promotes tumor growth and alters mitochondrial fitness in fibrolamellar carcinoma

Fibrolamellar carcinoma (FLC) is a rare liver cancer that disproportionately affects adolescents and young adults. Currently, no standard of care is available and there remains a dire need for new therapeutics. Most patients harbor the fusion oncogene DNAJB1-PRKACA (DP fusion), but clinical inhibitors are not yet developed and it is critical to identify downstream mediators of FLC pathogenesis. Here, we identify long noncoding RNA LINC00473 among the most highly upregulated genes in FLC tumors and determine that it is strongly suppressed by RNAi-mediated inhibition of the DP fusion in FLC tumor epithelial cells. We show by loss- and gain-of-function studies that LINC00473 suppresses apoptosis, increases the expression of FLC marker genes, and promotes FLC growth in cell-based and in vivo disease models. Mechanistically, LINC00473 plays an important role in promoting glycolysis and altering mitochondrial activity. Specifically, LINC00473 knockdown leads to increased spare respiratory capacity, which indicates mitochondrial fitness. Overall, we propose that LINC00473 could be a viable target for this devastating disease.

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2023-07-03 | The mechanism of LSM2 in the progression of live hepatocellular carcinoma was analyzed based on bioinformatics

Abstract Comprehensive analysis the expression and probable function of LSM2 in Live hepatocellular carcinoma (LIHC), and validation via in vitro experiments. Integrated use of database resources like TIMER2.0, GEPIA, LinkedOmics to examine the differential expression, survival prognosis, clinical pathological features, and functional enrichment of LSM2 in LIHC. The expression level of LSM2 in LIHC tissues and adjacent tissues was proven via immunohistochemical staining. Employing shRNA interference techniques to reduce the expression level of LSM2 in LIHC. The biological function of LSM2 in LIHC was detected by cell proliferation, cell cloning, cell scratch, cell migration, and invasion experiments in vitro. TIMER2.0 and GEPIA indicated that LSM2 was highly expressed in various cancers and was strongly associated with survival rates in patients with LIHC, cholangiocarcinoma, breast cancer, and renal clear cell carcinoma. LSM2 was highly expressed in LIHC, which was closely associated to the clinicopathological characteristics of patients, and the overall survival rate and disease-free survival rate of patients with high expression of LSM2 were lower than those with low expression of LSM2. Functional enrichment results revealed that LSM2 was involved to ribosome formation, DNA replication, cell cycle, metabolic processes, JAK-STAT signaling pathways, and FoxO signaling pathways. Compared with the paracancerous tissues, the expression of LSM2 in LIHC tissues was higher. Knockdown of LSM2 inhibited the proliferation, migration, and invasion of LIHC cells in in vitro experiments. LSM2 was highly expressed in LIHC and was related to a poor prognosis. Knockdown of LSM2 could inhibit the proliferation, migration, and invasion of LIHC cells.

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2023-01-05 | Oncogenic Addiction of Fibrolamellar Hepatocellular Carcinoma to the Fusion Kinase DNAJB1-PRKACA.

Gene fusions are drivers of many pediatric tumors. In fibrolamellar hepatocellular carcinoma (FLC), a fusion of DNAJB1 and PRKACA is the dominant recurrent mutation. Expression of the DNAJB1-PRKACA fusion gene in mice results in a tumor that recapitulates FLC. However, it is not known whether transient expression of DNAJB1-PRKACA is sufficient only to trigger tumor formation or whether ongoing expression is necessary for maintenance and progression. We screened short hairpin RNAs (shRNA) tiled over the fusion junction and identified several potent and specific candidates in vitro and two independent FLC patient-derived xenografts (PDX). We show that continued DNAJB1-PRKACA expression is not only required for continued tumor growth, but additionally its inhibition results in cell death. Inhibition of DNAJB1-PRKACA by an inducible shRNA in cells of PDX of FLC resulted in cell death in vitro. Induction of the shRNA inhibits FLC tumors growing in mice with no effect on xenografts from a hepatocellular carcinoma cell line engineered to express DNAJB1-PRKACA. Our results validate DNAJB1-PRKACA as the oncogene in FLC and demonstrate both a continued requirement for the oncogene for tumor growth as well as an oncogenic addiction that can be exploited for targeted therapies. We anticipate our approach will be useful for investigations of other fusion genes in pediatric cancers and spur development of precision therapies.

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2022-06-09 | Multiomic analysis of microRNA-mediated regulation reveals a proliferative axis involving miR-10b in fibrolamellar carcinoma.

Fibrolamellar carcinoma (FLC) is an aggressive liver cancer primarily afflicting adolescents and young adults. Most patients with FLC harbor a heterozygous deletion on chromosome 19 that leads to the oncogenic gene fusion, DNAJB1-PRKACA. There are currently no effective therapeutics for FLC. To address that, it is critical to gain deeper mechanistic insight into FLC pathogenesis. We assembled a large sample set of FLC and nonmalignant liver tissue (n = 52) and performed integrative multiomic analysis. Specifically, we carried out small RNA sequencing to define altered microRNA expression patterns in tumor samples and then coupled this analysis with RNA sequencing and chromatin run-on sequencing data to identify candidate master microRNA regulators of gene expression in FLC. We also evaluated the relationship between DNAJB1-PRKACA and microRNAs of interest in several human and mouse cell models. Finally, we performed loss-of-function experiments for a specific microRNA in cells established from a patient-derived xenograft (PDX) model. We identified miR-10b-5p as the top candidate pro-proliferative microRNA in FLC. In multiple human cell models, overexpression of DNAJB1-PRKACA led to significant upregulation of miR-10b-5p. Inhibition of miR-10b in PDX-derived cells increased the expression of several potentially novel target genes, concomitant with a significant reduction in metabolic activity, proliferation, and anchorage-independent growth. This study highlights a potentially novel proliferative axis in FLC and provides a rich resource for further investigation of FLC etiology.

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2020-04-06 | Immune prognostic implications of PSMD14 and its associated genes signatures in hepatocellular carcinoma

Abstract Background PSMD14 played a vital roles initiation and progression of hepatocellular carcinoma (HCC). However, PSMD14 and its-related genes for the immune prognostic implications of HCC patients have rarely been analyzed. Therefore, we aimed to explore gene signatures and immune prognostic values of PSMD14 and its-related genes in HCC. Method Analyzed the expression of PSMD14 in multiple databases, and clinicopathologic characteristics associated with PSMD14 overall survival using Wilcoxon signed-ranktest, logistic and Cox regression, Kaplan-Meier method. An immune prognostic signature (including RBM45, PSMD1, OLA1, CCT6A, LCAT and IVD) was constructed and validated using the co-expression and cox regression analyses in TCGA, ICGC and TIMER datasets. Gene Set Enrichment Analysis (GSEA) was performed using TCGA data set. Results Increased PSMD14 expression in HCC was significantly associated with poor prognosis and clinicopathologic characteristics (grade, histologic stage, surgical approach and T stage, all p-values < 0.05). A total of six PSMD14-related genes were detected, which markedly related to overall survival and immune infiltrating levels in HCC patients. Using cox regression analysis, the PSMD14 and its-related genes were found to be an independent prognostic factor for HCC survival. Calibration curves confirmed good consistency between clinical nomogram prediction and actual observation. Immune prognostic model suggests that patients in the high‐risk group shown significantly poorer survival than patients in the low‐risk group. Conclusion We screened potential immune prognostic genes and constructed and verified a novel PSMD14-based prognostic model of HCC, which provides new potential prognostic biomarkers and therapeutic targets and lays a theoretical foundation for immunotherapy of HCC.

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2026-08-13 | Management of rare primary liver cancers: combined hepato-cholangiocarcinoma and fibrolamellar hepatocellular carcinoma.

Fibrolamellar hepatocellular carcinoma (FLC) and combined hepatocellular-cholangiocarcinoma (cHCC-CCA) are rare primary liver cancers for which limited evidence is available to guide clinical management. cHCC-CCA could arises in the context of chronic liver disease and shares epidemiological and molecular features with both hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (iCCA), reflecting its intermediate biological and clinical position between these two entities. Its diagnosis remains challenging particularly when based on tumor biopsy specimens. Surgical resection remains the standard of care when feasible. Liver transplantation, trans-arterial chemoembolization, ablation, or selective internal radiation therapy have been used in selected cases. For advanced disease, systemic treatments commonly used for HCC or CCA have been applied in real-world practice, including platinum-based chemotherapy, tyrosine kinase inhibitors, and more recently immunotherapy. In addition, molecularly guided therapies have been explored, as targetable genetic alterations can be identified in a subset of cHCC-CCA. In contrast, FLC typically arises in non-cirrhotic livers of young patients and is frequently associated with metastatic disease, including lymph node involvement. At the molecular level, FLC is characterized by the presence of the DNAJB1-PRKACA fusion and by elevated serum procalcitonin secreted by the tumor. Surgical resection remains the cornerstone of treatment when feasible, but data guiding systemic therapy in unresectable cases are limited. Several regimens, including gemcitabine-oxaliplatin, lenvatinib, and immunotherapy, have been evaluated. More recently, clinical trials have reported antitumor activity with combinations of immunotherapy and vaccines targeting the DNAJB1-PRKACA fusion. To improve patient care, since cHCC-CCA and FLC are rare subtypes of liver cancer, we need to compile prospective samples and clinical data from the patients, including their response to any type of treatment at the national and international levels.

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2026-05-28 | FusionVAC22_02, a phase I clinical trial in progress: Adjuvant DNAJB1-PRKACA fusion transcript–based peptide T-cell activator for fibrolamellar hepatocellular carcinoma (FLC) and other tumor entities carrying the oncogenic driver fusion.

TPS4270 Background: The DNAJB1-PRKACA fusion transcript was detected as the oncogenic driver of tumor pathogenesis in fibrolamellar hepatocellular carcinoma (FLC) and other cancers, e.g. oncocytic neoplasms of pancreas and bile duct. We and others have shown that the fusion protein can be targeted by T cell-based immunotherapy: Application of Fusion-VAC-XS15 a peptide-based T cell activator including the TLR1/2 agonist XS15 emulsified in Montanide ISA 51 VG in two FLC patients was well tolerated without systemic side effects and induced long-lasting T-cell response accompanied by disease remission with a progression-free survival of up to 80 months and 60 months in both patients (Bauer et al. Nat. Commun., 2022). Based on these promising data, Fusion-VAC-XS15 combined with Atezolizumab is currently under evaluation in the advanced or metastatic situation since October 2023 (Hackenbruch et al. Front Oncol., 2024; NCT05937295). For localized FLC, surgical resection still represents the only curative treatment option but shows high relapse rates which underscores the high medical need for adjuvant treatment options. We here present the FusionVAC22_02 trial, which evaluates Fusion-VAC-XS15 as an adjuvant therapeutic in FLC patients who have reached complete remission. Methods: FusionVAC22_02 is a Phase I open label, multicentric clinical trial evaluating immunogenicity along with safety, toxicity and first signs of clinical efficacy of Fusion-VAC-XS15 as adjuvant treatment, in 20 patients with FLC or other cancers with proven DNAJB1-PRKACA fusion protein, and lacking adjuvant treatment options. One key eligibility criterion is achievement of complete remission (e.g. due to surgery, radiotherapy, local intervention or systemic treatment) according to RECIST1.1. Of note, a history of liver transplantation or prior immune-mediated side effects (e.g. after treatment with checkpoint-inhibitors) are no exclusion criteria. Fusion-VAC-XS15 is applied twice in a 4-week interval, with an optional booster 56 days after the second application, followed by a 6-month follow-up. Primary objectives include assessment of immunogenicity in terms of peptide-specific T cell responses, as well as evaluation of safety and toxicity. Safety assessment is based on the frequency of adverse events according to CTCAE v5.0. Clinical efficacy is determined by RECIST1.1 assessment on imaging. Recruitment started in July 2025, nine FLC patients from various parts of the world have been included and treated so far, four of which have reached the primary endpoint. Clinical trial information: NCT06789198 .

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2026-05-28 | Survival in patients with hepatobiliary and pancreatic cancers taking GLP-1 receptor agonist therapy.

e16276 Background: Metabolic syndrome has been identified as an independent risk factor for hepatobiliary and pancreatic cancers. Although it is linked as a causative factor, the impact of treatment directed towards risk-reduction and the result on oncologic outcomes remain limited. Since the introduction of glucagon-like peptide-1 receptor agonists (GLP1), emerging evidence has shown associations with reducing the incidence of several cancers, including endometrial, ovarian and meningioma. To our knowledge, this is the first large-scale study to specifically address survival outcomes in patients with hepatobiliary and pancreatic cancers taking GLP1. Methods: A retrospective cohort study using the TriNetX Research Network from 158 healthcare organizations (HCO) from the Global Collaborative Network was carried out. We identified patients who were diagnosed with hepatocellular carcinoma (HCC), fibrolamellar HCC, cholangiocarcinoma, gallbladder and pancreatic cancer using the ICD-10 code and either received GLP1 treatment or not. Cohorts were balanced using propensity score matching for age at diagnosis, female, Black race, body mass index, comorbidities, and concomitant medications. Kaplan-Meier survival curves were used to assess overall survival (OS). Results: From 01/01/2010-12/31/2025, a total of 337,275 patients who had a hepatobiliary or pancreatic cancer were identified from 38 HCO, of these 4,404 patients started GLP1 after the diagnosis and 332,871 were not on GLP1. After balancing, each cohort had a total of 3,607 patients. Patients using GLP1 were noted to be younger and with more diverse ethnical and racial background. A higher body mass index was reported in those patients taking GLP1. Statistically significant differences were noted in laboratory values showing a lower creatinine, ALT and AST in those taking GLP1. Patients who were diagnosed with a hepatobiliary or pancreatic cancer who were taking GLP1 were noted to have better OS than those who did not [p-value < 0.001]. At 10 years, the median survival for those who received GLP1 was not reached vs 1,133 days for those who didn’t. Survival probabilities for patients using GLP1 vs. not at 6, 12, and 24 months were 92% vs. 77%, 86% vs. 67%, and 80% vs. 56%, respectively. Conclusions: Patients with hepatobiliary and pancreatic malignancies receiving GLP1 therapy demonstrated superior survival outcomes. This survival benefit may be driven by the anti-inflammatory properties of GLP1, mediated through both immunomodulatory pathways and the direct attenuation of oncogenic signaling and proliferation. Beyond gastrointestinal malignancies, emerging evidence suggests that GLP1 exert an effect against various metabolically driven malignancies. Given the expanding clinical indications for GLP1 therapy, further investigation into its broader oncologic effect is warranted.

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2025-11-04 | A Pilot Study of a DNAJB1-PRKACA Fusion Kinase Peptide Vaccine Combined With Nivolumab and Ipilimumab for Patients With Fibrolamellar Hepatocellular Carcinoma

The primary objective of the trial is the safety and tolerability of administering a vaccine targeting the DNAJB1-PRKACA fusion kinase, in combination with nivolumab and ipilimumab in patients with unresectable or metastatic FLC and with non-FLC solid tumors and to assess the T-cell response.

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2024-05-09 | Clinical significance of PNO1 as a novel biomarker and therapeutic target of hepatocellular carcinoma.

The RNA-binding protein PNO1 plays an essential role in ribosome biogenesis. Recent studies have shown that it is involved in tumorigenesis; however, its role in hepatocellular carcinoma (HCC) is not well understood. The purpose of this study was to examine whether PNO1 can be used as a biomarker of HCC and also examine the therapeutic potential of PNO1 knockout for the treatment of HCC. PNO1 expression was upregulated in HCC and associated with poor prognosis. PNO1 expression was positively associated with tumour stage, lymph node metastasis and poor survival. PNO1 expression was significantly higher in HCC compared to that in fibrolamellar carcinoma or normal tissues. Furthermore, HCC tissues with mutant Tp53 expressed higher PNO1 than those with wild-type Tp53. PNO1 knockout suppressed cell viability, colony formation and EMT of HCC cells. Since activation of Notch signalling pathway promotes HCC, we measured the effects of PNO1 knockout on the components of Notch pathway and its targets. PNO1 knockout suppressed Notch signalling by modulating the expression of Notch ligands and their receptors, and downstream targets. PNO1 knockout also inhibited genes involved in surface adhesion, cell cycle, inflammation and chemotaxis. PNO1 knockout also inhibited colony and spheroid formation, cell migration and invasion, and markers of stem cells, pluripotency and EMT in CSCs. Overall, our data suggest that PNO1 can be used as a diagnostic and prognostic biomarker of HCC, and knockout of PNO1 by CRISPR/Cas9 can be beneficial for the management of HCC by targeting CSCs.

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