RARE DISEASE
Mucopolysaccharidosis type 10
Mucopolysaccharidosis type 10
Mucopolysaccharidosis type 10
Synonyms: Mucopolysaccharidosis type X, MSP type X,MPS10 Mucopolysaccharidosis due to ARSK deficiency
Synonyms: Mucopolysaccharidosis type X, MSP type X,MPS10 Mucopolysaccharidosis due to ARSK deficiency
Synonyms: Mucopolysaccharidosis type X, MSP type X,MPS10 Mucopolysaccharidosis due to ARSK deficiency
Drug discovery
133
papers
On emerging therapies
Overview
Mucopolysaccharidosis type 10 (MPS X) is an ultra-rare autosomal recessive lysosomal storage disorder caused by deficient arylsulfatase K (ARSK) activity, leading to glycosaminoglycan accumulation. Clinical features include disproportionate short stature, skeletal dysplasia, coarse facial features, hearing loss, and potential cardiac/liver involvement [1][19]. Cognitive impairment is typically absent. Diagnosis involves enzyme assays and genetic testing for ARSK mutations [19].
Therapies
Supportive care: Surgical correction of skeletal abnormalities, hearing aids, and cardiac monitoring [19]
No approved disease-modifying therapies; enzyme replacement therapy (ERT) and hematopoietic stem cell transplantation (HSCT) remain investigational [16][20]
Preclinical studies exploring gene therapy approaches [20]
Categories: rare bone diseases, rare developmental anomalies during embryogenesis, rare genetic diseases, rare inborn errors of metabolism, rare neurological diseases, rare ophthalmic disorders, rare transplant-related disorders
Research Papers
133 drug discovery papers about Mucopolysaccharidosis type 10, with 4 first-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
133 drug discovery papers about Mucopolysaccharidosis type 10, with 4 first-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
proteins
2023-10-29 | The Experiences and Challenges of Mothers of Adult Patients with Mucopolysaccharidosis; Mothers of Adults with Severe Mucopolysaccharidosis
Although many studies have examined the treatment and symptoms of children with MPS, very few studies have focused on adults with MPS. This study aimed to clarify the experiences and challenges of mothers of adult patients with severe mucopolysaccharidosis (MPS) who live at home. The data from semi-structured interviews with twelve mothers (patients’ ages ranged 19-44 years) were analyzed. The following six categories were extracted: fear that the disease will continue to progress despite treatment, tired of shifting gears in care and healing, fatigue due to multi-department visits to control the progression of the disease, the pain of not being able to see a bright future, desperate for new treatment methods for MPS, and developing care and a place to stay after parental support is gone. A unique issue of patients with MPS is that enzyme replacement therapy must be continued throughout life, and there is an urgent need to create a system that allows patients with severe MPS to receive enzyme replacement therapy for the rest of their lives, even after their parents pass away.
2023-06-27 | Hydrogel Delivery Device for the In Vitro and In Vivo Sustained Release of Active rhGALNS Enzyme
Morquio A disease is a genetic disorder resulting in N-acetylgalactosamine-6-sulfate sulfatase (GALNS) deficiency, and patients are currently treated with enzyme replacement therapy via weekly intravenous enzyme infusions. A means of sustained enzyme delivery could improve patient quality of life by reducing the administration time, frequency of hospital visits, and treatment cost. In this study, we investigated poly(ethylene-glycol) (PEG) hydrogels as a tunable, hydrolytically degradable drug delivery system for the encapsulation and sustained release of recombinant human GALNS (rhGALNS). We evaluated hydrogel formulations that optimized hydrogel gelation and degradation time while retaining rhGALNS activity and sustaining rhGALNS release. We observed the release of active rhGALNS for up to 28 days in vitro from the optimized formulation. rhGALNS activity was preserved in the hydrogel relative to buffer over the release window, and encapsulation was found to have no impact on the rhGALNS structure when measured by intrinsic fluorescence, circular dichroism, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In vivo, we monitored the retention of fluorescently labeled rhGALNS in C57BL/6 albino mice when administered via subcutaneous injection and observed rhGALNS present for up to 20 days when delivered in a hydrogel versus 7 days in the buffer control. These results indicate that PEG hydrogels are suitable for the encapsulation, preservation, and sustained release of recombinant enzymes and may present an alternative method of delivering enzyme replacement therapies that improve patient quality of life.
2023-02-24 | A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution
Currently available enzyme replacement therapies for lysosomal storage diseases are limited in their effectiveness due in part to short circulation times and suboptimal biodistribution of the therapeutic enzymes. We previously engineered Chinese hamster ovary (CHO) cells to produce α-galactosidase A (GLA) with various N-glycan structures and demonstrated that elimination of mannose-6-phosphate (M6P) and conversion to homogeneous sialylated N-glycans prolonged circulation time and improved biodistribution of the enzyme following a single-dose infusion into Fabry mice. Here, we confirmed these findings using repeated infusions of the glycoengineered GLA into Fabry mice and further tested whether this glycoengineering approach, Long-Acting-GlycoDesign (LAGD), could be implemented on other lysosomal enzymes. LAGD-engineered CHO cells stably expressing a panel of lysosomal enzymes [aspartylglucosamine (AGA), beta-glucuronidase (GUSB), cathepsin D (CTSD), tripeptidyl peptidase (TPP1), alpha-glucosidase (GAA) or iduronate 2-sulfatase (IDS)] successfully converted all M6P-containing N-glycans to complex sialylated N-glycans. The resulting homogenous glycodesigns enabled glycoprotein profiling by native mass spectrometry. Notably, LAGD extended the plasma half-life of all three enzymes tested (GLA, GUSB, AGA) in wildtype mice. LAGD may be widely applicable to lysosomal replacement enzymes to improve their circulatory stability and therapeutic efficacy.
2022-03-11 | Improving yield of a recombinant biologic in a Brassica hairy root manufacturing process
Hairy root systems have proven to be a viable alternative for recombinant protein production. For recalcitrant proteins, maximizing the productivity of hairy root cultures is essential. The aim of this study was to optimize a Brassica rapa rapa hairy root process for secretion of α-L-iduronidase (IDUA), a biologic of medical value. The process was first optimized with hairy roots expressing eGFP. For the biomass optimization, the highest biomass yields were achieved in modified Gamborg B5 culture medium. For the secretion induction, the optimized secretion media was obtained with additives (1.5g/l PVP + 1mg/l 2,4-D + 20.5g/l KNO ) resulting in 3.4 fold eGFP secretion when compared to the non-induced control. These optimized conditions were applied to the IDUA-expressing hairy root clone, confirming that the highest yields of secreted IDUA occurred when using the already defined additive combination. The functionality of the IDUA protein, secreted and intracellular, was confirmed with an enzymatic activity assay. A >150-fold increase of the IDUA activity was observed using an optimized secretion medium, compared with a non-induced medium. We have proven that our B. rapa rapa hairy root system can be harnessed to secrete recalcitrant proteins, illustrating the high potential of hairy roots in plant molecular farming.
2021-11-19 | Elosulfase Alfa Treatment in Morquio A Patients in Iran: A Before and After Study
Background: Morquio A, an autosomal recessive lysosomal storage disease, is caused by a defect in the enzyme N-acetyl-galactosamine-6-sulfatase. This leads to the accumulation of the glycosaminoglycans chondroitin-6-sulfate (C6S) and keratan sulfate (KS), resulting in various skeletal manifestations, multisystemic impairments, and significant morbidities. Objectives: This study aimed to evaluate the impact of the addition of elosulfase alfa to the hospital protocol on treating Iranian pediatricians with Morquio A syndrome. Methods: A before and after study was conducted on ten patients with Morquio A syndrome diagnosis from 2019 to 2020. Elosulfase alfa was prescribed with the standard dose of 2 mg/kg/weekly IV infusion for 54 weeks. Then, growth indices, quality of life, and cardiopulmonary data were collected by research assistants using a pre-designed check. Data were entered in SPSS version 23. Quantitative variables were compared between the two periods using the Student's t-test, and qualitative variables were compared using the χ2 test or Fisher's exact test. Results: Ten pediatricians with MPS IV were included, seven of whom were female, with the mean age of 5.8 ± 2.3 years. The meantime for walking 6 m (P = 0.005), standing (P = 0.005), the stair climb test (P = 0.007), and quality of life (P = 0.015) had significant statistical difference before and after treatment by elosulfase alfa. Conclusions: The addition of elosulfase alfa in patients with Morquio A syndrome was associated with a significant improvement outcome in the ‘after’ treatment period.
small molecules
2024-03-28 | Investigation of the Oxidative Process by Measuring Total Antioxidant Capacity and Total Oxidant Capacity in Patients with Mucopolysaccharidosis: Cross-Sectional Prospective Study
Objective: To compare the oxidative stress status between the patients with mucopolysaccharidosis who received and did not receive enzyme replacement therapy and healthy control group. Material and Methods: A retrospective study of mucopolysaccharidosis patients with analysis of the total oxidant and antioxidant capacity levels. Results: We included 29 mucopolysaccharidosis patients aged between 1-18 years and 50 healthy children. Total antioxidant capacity and total oxidant capacity parameters between this patients and healthy control group were compared. The median age of the patients who were included in the study was 9.0 years and the median age of the cases included in the control group was 8.0 years. Total antioxidant capacity, total oxidant capacity and Oxidative Stress Index (OSI) values did not differ significantly in patients receiving enzyme replacement therapy compared to patients who did not receive enzyme replacement therapy. A comparison of the oxidant and antioxidant system parameters between the patient and control groups revealed a distinct difference, with the control group exhibiting a considerably higher total antioxidant capacity than mucopolysaccharidosis patients, and the patient group showing a significantly higher total oxidant capacity and OSI. Conclusion: In this study, it was observed that the antioxidant defence system decreased in patients with mucopolysaccharidoses. It is thought that, in addition to standard treatments, the administration of antioxidant treatments and supporting nutrition in mucopolysaccharidosis patients will increase the quality of life of the patients. Regular sleep, eating healthy foods and doing regular exercise would also increase the effectiveness of these supplements.
2023-11-17 | Gold nanoparticles decorated with monosaccharides and sulfated ligands as potential modulators of the lysosomal enzyme N-acetylgalactosamine-6-sulfatase (GALNS).
N-Acetylgalactosamine-6-sulfatase (GALNS) is an enzyme whose deficiency is related to the lysosomal storage disease Morquio A. For the development of effective therapeutic approaches against this disease, the design of suitable enzyme enhancers (i.e. pharmacological chaperones) is fundamental. The natural substrates of GALNS are the glycosaminoglycans keratan sulfate and chondroitin 6-sulfate, which mainly display repeating units of sulfated carbohydrates. With a biomimetic approach, gold nanoparticles (AuNPs) decorated with simple monosaccharides, sulfated ligands (homoligand AuNPs), or both monosaccharides and sulfated ligands (mixed-ligand AuNPs) were designed here as multivalent inhibitors of GALNS. Among the homoligand AuNPs, the most effective inhibitors of GALNS activity are the β-D-galactoside-coated AuNPs. In the case of mixed-ligand AuNPs, β-D-galactosides/sulfated ligands do not show better inhibition than the β-D-galactoside-coated AuNPs. However, a synergistic effect is observed for α-D-mannosides in a mixed-ligand coating with sulfated ligands that reduced IC50 by one order of magnitude with respect to the homoligand α-D-mannoside-coated AuNPs. SAXS experiments corroborated the association of GALNS with β-D-galactoside AuNPs. These AuNPs are able to restore the enzyme activity by almost 2-fold after thermal denaturation, indicating a potential chaperoning activity towards GALNS. This information could be exploited for future development of nanomedicines for Morquio A. The recent implications of GALNS in cancer and neuropathic pain make these kinds of multivalent bionanomaterials of great interest towards multiple therapies.
2023-08-31 | Synthesis and Evaluation of Novel Triaryl Derivatives with Readthrough-Inducing Activity
The readthrough mechanism, which skips the premature termination codon and restores the biosynthesis of the defective enzyme, is an emerging therapeutic tactic for nonsense mutation-related diseases, such as Hurler syndrome, a type of mucopolysaccharidosis. In the present study, novel triaryl derivatives were synthesized and their readthrough-inducing activities were evaluated by a luciferase reporter assay with a partial α-L-iduronidase (IDUA) DNA sequence containing the Q70X nonsense mutation found in Hurler syndrome and by measuring the enzyme activity of IDUA knockout cells transfected with the mutant IDUA gene. KY-516, a representative compound in which the meta position carboxyl group of the left ring of the clinically used ataluren was converted to the para position sulfamoylamino group, the central ring to triazole, and the right ring to cyanobenzene, exhibited the most potent readthrough-inducing activity in the Q70X/luciferase reporter assay. In Q70X mutant IDUA transgenic cells, KY-516 significantly increased enzyme activity at 0.1 µM. After the oral administration of KY-516 (10 mg/kg), the highest plasma concentration of KY-516 was above 5 µM in rats. These results indicate that KY-516, a novel triaryl derivative, exhibits potent readthrough-inducing activity and has potential as a therapeutic agent for Hurler syndrome.
2020-12-11 | Molecular Insights into Determinants of Translational Readthrough and Implications for Nonsense Suppression Approaches
The fidelity of protein synthesis, a process shaped by several mechanisms involving specialized ribosome regions and external factors, ensures the precise reading of sense and stop codons. However, premature termination codons (PTCs) arising from mutations may, at low frequency, be misrecognized and result in PTC suppression, named ribosome readthrough, with production of full-length proteins through the insertion of a subset of amino acids. Since some drugs have been identified as readthrough inducers, this fidelity drawback has been explored as a therapeutic approach in several models of human diseases caused by nonsense mutations. Here, we focus on the mechanisms driving translation in normal and aberrant conditions, the potential fates of mRNA in the presence of a PTC, as well as on the results obtained in the research of efficient readthrough-inducing compounds. In particular, we describe the molecular determinants shaping the outcome of readthrough, namely the nucleotide and protein context, with the latter being pivotal to produce functional full-length proteins. Through the interpretation of experimental and mechanistic findings, mainly obtained in lysosomal and coagulation disorders, we also propose a scenario of potential readthrough-favorable features to achieve relevant rescue profiles, representing the main issue for the potential translatability of readthrough as a therapeutic strategy.
2020-07-31 | Screening, Synthesis, and Evaluation of Novel Isoflavone Derivatives as Inhibitors of Human Golgi β-Galactosidase
The genes GLB1 and GALC encode GLB1 isoform 1 and galactocerebrosidase, respectively, which exhibit β-galactosidase activity in human lysosomes. GLB1 isoform 1 has been reported to play roles in rare lysosomal storage diseases. Further, its β-galactosidase activity is the most widely used biomarker of senescent and aging cells; hence, it is called senescence-associated β-galactosidase. Galactocerebrosidase plays roles in Krabbe disease. We previously reported a novel β-galactosidase activity in the Golgi apparatus of human cells; however, the protein responsible for this activity could not be identified. Inhibitor-derived chemical probes can serve as powerful tools to identify the responsible protein. In this study, we first constructed a cell-based high-throughput screening (HTS) system for Golgi β-galactosidase inhibitors, and then screened inhibitors from two compound libraries using the HTS system, in vitro assay, and cytotoxicity assay. An isoflavone derivative was identified among the final Golgi β-galactosidase inhibitor compound hits. Molecular docking simulations were performed to redesign the isoflavone derivative into a more potent inhibitor, and six designed derivatives were then synthesized. One of the derivatives, ARM07, exhibited potent inhibitory activity against β-galactosidase, with an IC50 value of 14.8 µM and competitive inhibition with Ki value of 13.3 µM. Furthermore, the in vitro and cellular inhibitory activities of ARM07 exceeded those of deoxygalactonojirimycin. ARM07 may contribute to the development of affinity-based chemical probes to identify the protein responsible for the newly discovered Golgi β-galactosidase activity. The therapeutic relevance of ARM07 against lysosomal storage diseases and its effect on senescent cells should be evaluated further.
cell therapies
2022-12-25 | Modifying enzyme replacement therapy – A perspective
Abstract Several diseases are caused by the lack of functional proteins, including lysosomal storage diseases or haemophilia A and B. Patients suffering from one of these diseases are treated via enzyme replacement therapies to restore the missing protein. Although this treatment strategy prevents some disease symptoms, enzyme replacement therapies are very expensive and require very frequent infusions, which can cause infusion adverse reactions and massively impair the quality of life of the patients. This review proposes a technology to sustainably produce proteins within the patient to potentially make frequent protein‐infusions redundant. This technology is based on blood circulating immune cells as producers of the needed therapeutic protein. To ensure a stable protein concentration over time the cells are equipped with a system, which induces cell proliferation when low therapeutic protein levels are detected and a system inhibiting cell proliferation when high therapeutic protein levels are detected.
2019-06-18 | A decade of excellent transplant survival in children with inherited metabolic diseases: A report from a single metabolic transplant centre in Europe
Hematopoietic cell transplantation (HCT) confers a long-term disease-modifying therapy for transplant-permissive inherited metabolic diseases (IMDs). We examined the overall survival (OS) and engrafted survival (ES) of children with IMDs, who received first HCT at Royal Manchester Children's hospital from 1985 to 2016. A total of 137 children with IMDs were included in this analysis (historical cohort [1985-2006], n＝65; current cohort [2007-2016], n＝72). Primary diagnoses included mucopolysaccharidoses (81%), X-linked adrenoleukodystrophy (6%), metachromatic leukodystrophy (4%), mannosidosis (3%), Wolman disease (2%), and other conditions (4%). The five-year OS has increased from 65% (95% confidence interval [CI], 52%-76%) in the historical cohort to 91% (95% CI, 81%-96%) in the current cohort (P<0.001). Moreover, the five-year ES, which was 64% (95 CI%, 56%-72%) for the entire cohort, has doubled from 41% (95% CI, 29%-53%) in the historical cohort to 85% (95% CI, 75%-92%) in the current cohort (P<0.001). The proportion of patients with graft failure has decreased from 37% in the historical cohort to 8% in the current cohort (P<0.001). In patients who received a second transplant, 13 out of 20 patients (65%) in the historical cohort and all four in the current cohort were alive and engrafted. Of 82 survivors followed-up at Manchester, 80% and 20% had full and mixed chimerism, respectively. Although this study was restricted to a single center, our findings show that HCT is an increasingly safe procedure and provides long-lasting endogenous enzyme replacement therapy for children with IMDs in the modern era of HCT.
2019-02-01 | Preliminary results demonstrate engraftment with minimal neutropenia with MGTA-456, a CD34expanded cord blood (CB) product in patients transplanted for inherited metabolic disorders (IMD)
Mucopolysaccharidosis type IH (Hurler Syndrome), metachromatic leukodystrophy (MLD), globoid cell leukodystrophy (GLD) and cerebral adrenoleukodystrophy (cALD) are progressive diseases treatable with allogeneic hematopoietic stem cell transplantation (HSCT). While cord blood is a common stem cell source, it may be associated with prolonged neutropenia and graft failure. MGTA-456 is a cell therapy produced from a single CB unit using an aryl hydrocarbon receptor antagonist in a 15-day expansion culture of CD34+cells. In previous studies, patients with hematologic malignancies demonstrated a median 324-fold expansion of CD34+cells, and the time to neutrophil recovery was significantly reduced by a median of 9 days compared to historical controls. As higher CD34+doses have been correlated with improved engraftment and outcomes in inherited metabolic disease patients, we initiated a Phase 2, open-label trial to enroll up to 12 patients with a diagnosis of MPS1, cALD, MLD, and GLD utilizing CB units matched at ≥6 of 8 HLA loci using allele-based typing. The HSCT regimen consists of anti-thymocyte globulin (days -9 to -6), fludarabine (days -5 to -2) and busulfan (days -5 to -2), with cyclosporin and methylprednisolone as graft vs. host disease prophylaxis. In four patients treated thus far, a marked expansion of CD34+cells (median 482-fold) was observed. Two patients had no days of neutropenia and 2 patients had 1 and 4 days respectively, in contrast to a mean of 7.8 days in a historical Minnesota cohort (N=27) with identical conditioning. Donor myeloid chimerism (≥98%) was achieved by day +14 in all patients. One patient developed autoimmune cytopenia, a known complication unrelated to MSGT 456, resulting in death at day +143. Based on these promising data, MGTA-456 has potential to improve hematopoietic cell recovery and transplant-related outcomes in patients undergoing HSCT for patients with Inherited metabolic disorders.
2017-04-29 | Delivery of Cargo to Lysosomes Using GNeosomes
Liposomes have been used to improve the intracellular delivery of a variety of cargos. Encapsulation of cargos in liposomes leads to improved plasma half-lives and minimized degradation. Here, we present a method for improving the selective delivery of liposomes to the lysosomes using a guanidinylated neomycin (GNeo) transporter. The method for synthesizing GNeo-lipids, incorporating them into liposomes, and the enhanced lysosomal delivery of encapsulated cargo are presented. GNeo-liposomes, termed GNeosomes, are capable of delivering a fluorescent dye to the lysosomes of Chinese hamster ovary cells as shown using confocal microscopy. GNeosomes can also be used to deliver therapeutic quantities of lysosomal enzymes to fibroblasts isolated from patients with a lysosomal storage disorder.
2016-09-15 | Cell Therapy of Corneal Diseases
Mesenchymal stem cells isolated from connective tissues are pluripotent and differentiate into phenotypes of connective tissue cell lineages (osteoblasts, chondrocytes, and adipocytes) in vitro and in vivo. They have been used to treat mouse models of connective tissue disease such as lumican-null (Lum−/−) and mucopolysaccharidosis (Gusb) mice. Mesenchymal stem cells have unique immunosuppressive properties allowing evasion of host rejection; thus, they are valuable tools for cell therapy of congenital and acquired diseases involving immune dysfunction of multiple tissues including ocular surface tissues (cornea). We previously showed that human umbilical mesenchymal stem cells (UMSCs) modulated host immune responses, enabling them to survive xenograft transplantation. In vitro, UMSCs modulated inflammatory cells by inhibiting adhesion and invasion, and inducing cell death. UMSCs also regulated M1/M2 macrophage polarization and induced T-regulatory cell maturation from naive intraperitoneal cavity lavage cells. UMSCs exposed to inflammatory cells synthesized a rich extracellular glycocalyx composed of hyaluronan (HA) bound to the heavy chains (HCs) of inter-alpha-trypsin inhibitor (HC-HA), which contains tumor necrosis factor-α-stimulated gene 6 (TSG6) that catalyzes the transfer of HCs to HA, versican, and pentraxin-3. Our in vivo and in vitro results showed that the glycocalyx regulated inflammatory cells, allowing UMSCs to survive host immune rejection. Administration of antibodies against glycocalyx constituents or digestion with hyaluronidase and chondroitinase ABC abolished the UMSCs' ability to modulate immune responses. Treatment with anti-CD44 antibodies also diminished modulation of M2 macrophages by UMSCs, indicating that cell surface CD44 is required for correct UMSC glycocalyx assembly to modulate inflammatory cells.
gene therapies
2024-02-09 | REMOVER-PITCh: microhomology-assisted long-range gene replacement with highly multiplexed CRISPR-Cas9
A variety of CRISPR-Cas9-based gene editing technologies have been developed, including gene insertion and gene replacement, and applied to the study and treatment of diseases. While numerous studies have been conducted to improve the efficiency of gene insertion and to expand the system in various ways, there have been relatively few reports on gene replacement technology; therefore, further improvements are still needed in this context. Here, we developed the REMOVER-PITCh system to establish an efficient long-range gene replacement method and demonstrated its utility at two genomic loci in human cultured cells. REMOVER-PITCh depends on microhomology-assisted gene insertion technology called PITCh with highly multiplexed CRISPR-Cas9. First, we achieved gene replacement of about 20-kb GUSB locus using this system. Second, by applying the previously established knock-in-enhancing platform, the LoAD system, along with REMOVER-PITCh, we achieved the replacement of a longer gene region of about 200 kb at the ARSB locus. Our REMOVER-PITCh system will make it possible to remove and incorporate a variety of sequences from and into the genome, respectively, which will facilitate the generation of various disease and humanized models.
2023-05-27 | Etiology Understanding Is the Core of Skeletal Diagnosis
Background: The term idiopathic is a non-specific disease label, which fails account for proper etiological understanding. The frequent usage of the term idiopathic may indicate clinical incompetence. We describe a number of patients from our practice with a history of complex bone disorders who were falsely given the term idiopathic by other Institutes. The sole tool used by other Institutes to conclude osteoporosis as a diagnosis is the bone mineral density. Material and Methods: Two Austrian families and one unrelated adult female patient (total number of 12 subjects) have been referred to our department with referral letters confirming the diagnosis of idiopathic osteoporosis. All patients manifested a diverse constellation of skeletal deformities. In all these patients, regular courses of anti-resorptive treatment plus calcium and vitamin D supplements. These have been excessively prescribed. Results: Our diagnostic process which is based heavily on detailed clinical and radiological phenotypic characterizations, we refuted the diagnosis of idiopathic osteoporosis for all patients because of the clear-cut clinical features. For the first family, we suggested the diagnosis of a novel type of mucopolysaccharidosis (the genetic results confirmed the diagnosis of a novel subtype of mucopolysaccharidosis (through the identification of arylsulfatase K ARSK). For the second family which has been subjected to a lengthy useless investigation for almost more than a decade, we refuted the diagnosis of idiopathic osteoporosis and the whole exome sequencing was consistent with the phenotypic diagnosis Nasu-Hakola disease (heterozygous missense mutations in TYROBP gene). The other unrelated 42-year-old- female patient was correctly diagnosed with Nasu-Hakola disease. Conclusion: The objective of this study is to minimize and suppress the usage of the term idiopathic. It has been imperative to understand the sequence of pathological events that occurred in these families. The core for diagnosis is the etiological understanding, which stemmed from professional conscientiousness. Patients underwent misdiagnosis of idiopathic osteoporosis and were treated extensively with anti-resorptive agents, calcium supplements and vitamin D. Strikingly, these medications resulted in a combination of serious cranial and cerebral lesions because of massive pathological calcifications. Imaging analysis via conventional radiographs, MRI and CT scan showed massive calcifications of the cranial sutures, inter-clinoid ligaments and hyper-calcification of the skull base. Sadly speaking, early onset dementia emerged in connection with sclerosing leukoencephalopathy. We believe that the exogenous toxic effects of the huge amounts of bone supplements consumed by these patients enhanced the pre-existing metabolic bone and cerebral disorders. Our findings can ameliorate the etiological understanding of early onset dementia and other correlated cranial and skeletal lesions.
2019-03-01 | Enzymatic replacement therapy for lysosomal storage disorders: Drug evaluations review in Spain
In the European Union companies only need to demonstrate that the risk-benefit balance of the new drug is favourable to obtain the authorization to sell new drugs. Hence a comparison with available treatments, a cost-effectiveness analysis and the place in the therapy of the new drug are not required. Therefore, it is necessary to carry out these analyses in an additional study. In Spain there is no national government agency, which conducts a centralized evaluation and makes decisions on funding and are the Pharmacy and Therapeutics committees that take mandatory decisions for the entire region. To identify all drug assessments and health technology assessment reports of the enzymatic replacement treatments for the four LSD considered in this study, including Mucopolysaccharidosis (MPS), Gaucher, Fabry and Pompe disease at the national, regional and hospital level; and to summarize the efficacy in terms of outcome measures, adverse events, economic impact and final recommendations and indicate potential improvements. 19 reviews at the regional and hospital level and one therapeutic positioning report at national level for MPS were considered for this. In general, the drugs evaluated in LSD have proved to be effective in the most frequent clinical forms of disease. However, there are clinical phenotypes that still do not have available an effective treatment. It is expected that new therapies, such as intrathecal therapies or gene therapy could be evaluated in the different types of LSD demonstrating positive effects, especially in the previous phenotypes.
2019-02-03 | Donor-Free Gene Correction by Targeted Interhomolog Recombination
Spontaneous gene correction by interhomolog recombination (IHR) occasionally occurs to ameliorate genetic diseases of blood and skin 1–3 . Using an engineered endogenous gene as a reporter, we demonstrate that gene correction by IHR is normally infrequent (≤0.02%) but is stimulated by DSBs targeted by CRISPR/Cas9 to both homologous chromosomes; reaching frequencies of 0.5%. We further show that depletion of POLQ stimulates IHR frequencies 4-fold, to 2%, and promotes IHR in G2 phase, when recombination between replicated homologs can correct not only compound heterozygous but also autosomal dominant “gain-of-function” mutations, which present a special challenge for gene therapy. The strategies reported here will enable optimization of IHR for gene therapy in a variety of cell types. Advantages include the ability to correct gain-of-function mutations, no need for an exogenous donor, and the potential to limit damage to coding sequence by targeting IHR to introns.
2018-12-27 | Basic and Clinical Application of Adeno-Associated Virus–Mediated Genome Editing
Traditional gene therapy (gene replacement) has made a breakthrough in treating inherited diseases. Adeno-associated virus (AAV) has emerged as a highly promising vector with innate ability, boosting the development of gene replacement and gene targeting. With the recent advance of engineered nucleases that work efficiently in human cells, AAV mediated-genome editing with nucleases has raised hopes for in situ gene therapy of inherited and non-inherited diseases. Here, the applications of AAV-mediated genome editing are highlighted, and the prospect of AAV and nucleases that will render extension of such success in clinical gene therapy is discussed.
proteins
2023-10-29 | The Experiences and Challenges of Mothers of Adult Patients with Mucopolysaccharidosis; Mothers of Adults with Severe Mucopolysaccharidosis
Although many studies have examined the treatment and symptoms of children with MPS, very few studies have focused on adults with MPS. This study aimed to clarify the experiences and challenges of mothers of adult patients with severe mucopolysaccharidosis (MPS) who live at home. The data from semi-structured interviews with twelve mothers (patients’ ages ranged 19-44 years) were analyzed. The following six categories were extracted: fear that the disease will continue to progress despite treatment, tired of shifting gears in care and healing, fatigue due to multi-department visits to control the progression of the disease, the pain of not being able to see a bright future, desperate for new treatment methods for MPS, and developing care and a place to stay after parental support is gone. A unique issue of patients with MPS is that enzyme replacement therapy must be continued throughout life, and there is an urgent need to create a system that allows patients with severe MPS to receive enzyme replacement therapy for the rest of their lives, even after their parents pass away.
2023-06-27 | Hydrogel Delivery Device for the In Vitro and In Vivo Sustained Release of Active rhGALNS Enzyme
Morquio A disease is a genetic disorder resulting in N-acetylgalactosamine-6-sulfate sulfatase (GALNS) deficiency, and patients are currently treated with enzyme replacement therapy via weekly intravenous enzyme infusions. A means of sustained enzyme delivery could improve patient quality of life by reducing the administration time, frequency of hospital visits, and treatment cost. In this study, we investigated poly(ethylene-glycol) (PEG) hydrogels as a tunable, hydrolytically degradable drug delivery system for the encapsulation and sustained release of recombinant human GALNS (rhGALNS). We evaluated hydrogel formulations that optimized hydrogel gelation and degradation time while retaining rhGALNS activity and sustaining rhGALNS release. We observed the release of active rhGALNS for up to 28 days in vitro from the optimized formulation. rhGALNS activity was preserved in the hydrogel relative to buffer over the release window, and encapsulation was found to have no impact on the rhGALNS structure when measured by intrinsic fluorescence, circular dichroism, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In vivo, we monitored the retention of fluorescently labeled rhGALNS in C57BL/6 albino mice when administered via subcutaneous injection and observed rhGALNS present for up to 20 days when delivered in a hydrogel versus 7 days in the buffer control. These results indicate that PEG hydrogels are suitable for the encapsulation, preservation, and sustained release of recombinant enzymes and may present an alternative method of delivering enzyme replacement therapies that improve patient quality of life.
2023-02-24 | A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution
Currently available enzyme replacement therapies for lysosomal storage diseases are limited in their effectiveness due in part to short circulation times and suboptimal biodistribution of the therapeutic enzymes. We previously engineered Chinese hamster ovary (CHO) cells to produce α-galactosidase A (GLA) with various N-glycan structures and demonstrated that elimination of mannose-6-phosphate (M6P) and conversion to homogeneous sialylated N-glycans prolonged circulation time and improved biodistribution of the enzyme following a single-dose infusion into Fabry mice. Here, we confirmed these findings using repeated infusions of the glycoengineered GLA into Fabry mice and further tested whether this glycoengineering approach, Long-Acting-GlycoDesign (LAGD), could be implemented on other lysosomal enzymes. LAGD-engineered CHO cells stably expressing a panel of lysosomal enzymes [aspartylglucosamine (AGA), beta-glucuronidase (GUSB), cathepsin D (CTSD), tripeptidyl peptidase (TPP1), alpha-glucosidase (GAA) or iduronate 2-sulfatase (IDS)] successfully converted all M6P-containing N-glycans to complex sialylated N-glycans. The resulting homogenous glycodesigns enabled glycoprotein profiling by native mass spectrometry. Notably, LAGD extended the plasma half-life of all three enzymes tested (GLA, GUSB, AGA) in wildtype mice. LAGD may be widely applicable to lysosomal replacement enzymes to improve their circulatory stability and therapeutic efficacy.
2022-03-11 | Improving yield of a recombinant biologic in a Brassica hairy root manufacturing process
Hairy root systems have proven to be a viable alternative for recombinant protein production. For recalcitrant proteins, maximizing the productivity of hairy root cultures is essential. The aim of this study was to optimize a Brassica rapa rapa hairy root process for secretion of α-L-iduronidase (IDUA), a biologic of medical value. The process was first optimized with hairy roots expressing eGFP. For the biomass optimization, the highest biomass yields were achieved in modified Gamborg B5 culture medium. For the secretion induction, the optimized secretion media was obtained with additives (1.5g/l PVP + 1mg/l 2,4-D + 20.5g/l KNO ) resulting in 3.4 fold eGFP secretion when compared to the non-induced control. These optimized conditions were applied to the IDUA-expressing hairy root clone, confirming that the highest yields of secreted IDUA occurred when using the already defined additive combination. The functionality of the IDUA protein, secreted and intracellular, was confirmed with an enzymatic activity assay. A >150-fold increase of the IDUA activity was observed using an optimized secretion medium, compared with a non-induced medium. We have proven that our B. rapa rapa hairy root system can be harnessed to secrete recalcitrant proteins, illustrating the high potential of hairy roots in plant molecular farming.
2021-11-19 | Elosulfase Alfa Treatment in Morquio A Patients in Iran: A Before and After Study
Background: Morquio A, an autosomal recessive lysosomal storage disease, is caused by a defect in the enzyme N-acetyl-galactosamine-6-sulfatase. This leads to the accumulation of the glycosaminoglycans chondroitin-6-sulfate (C6S) and keratan sulfate (KS), resulting in various skeletal manifestations, multisystemic impairments, and significant morbidities. Objectives: This study aimed to evaluate the impact of the addition of elosulfase alfa to the hospital protocol on treating Iranian pediatricians with Morquio A syndrome. Methods: A before and after study was conducted on ten patients with Morquio A syndrome diagnosis from 2019 to 2020. Elosulfase alfa was prescribed with the standard dose of 2 mg/kg/weekly IV infusion for 54 weeks. Then, growth indices, quality of life, and cardiopulmonary data were collected by research assistants using a pre-designed check. Data were entered in SPSS version 23. Quantitative variables were compared between the two periods using the Student's t-test, and qualitative variables were compared using the χ2 test or Fisher's exact test. Results: Ten pediatricians with MPS IV were included, seven of whom were female, with the mean age of 5.8 ± 2.3 years. The meantime for walking 6 m (P = 0.005), standing (P = 0.005), the stair climb test (P = 0.007), and quality of life (P = 0.015) had significant statistical difference before and after treatment by elosulfase alfa. Conclusions: The addition of elosulfase alfa in patients with Morquio A syndrome was associated with a significant improvement outcome in the ‘after’ treatment period.
small molecules
2024-03-28 | Investigation of the Oxidative Process by Measuring Total Antioxidant Capacity and Total Oxidant Capacity in Patients with Mucopolysaccharidosis: Cross-Sectional Prospective Study
Objective: To compare the oxidative stress status between the patients with mucopolysaccharidosis who received and did not receive enzyme replacement therapy and healthy control group. Material and Methods: A retrospective study of mucopolysaccharidosis patients with analysis of the total oxidant and antioxidant capacity levels. Results: We included 29 mucopolysaccharidosis patients aged between 1-18 years and 50 healthy children. Total antioxidant capacity and total oxidant capacity parameters between this patients and healthy control group were compared. The median age of the patients who were included in the study was 9.0 years and the median age of the cases included in the control group was 8.0 years. Total antioxidant capacity, total oxidant capacity and Oxidative Stress Index (OSI) values did not differ significantly in patients receiving enzyme replacement therapy compared to patients who did not receive enzyme replacement therapy. A comparison of the oxidant and antioxidant system parameters between the patient and control groups revealed a distinct difference, with the control group exhibiting a considerably higher total antioxidant capacity than mucopolysaccharidosis patients, and the patient group showing a significantly higher total oxidant capacity and OSI. Conclusion: In this study, it was observed that the antioxidant defence system decreased in patients with mucopolysaccharidoses. It is thought that, in addition to standard treatments, the administration of antioxidant treatments and supporting nutrition in mucopolysaccharidosis patients will increase the quality of life of the patients. Regular sleep, eating healthy foods and doing regular exercise would also increase the effectiveness of these supplements.
2023-11-17 | Gold nanoparticles decorated with monosaccharides and sulfated ligands as potential modulators of the lysosomal enzyme N-acetylgalactosamine-6-sulfatase (GALNS).
N-Acetylgalactosamine-6-sulfatase (GALNS) is an enzyme whose deficiency is related to the lysosomal storage disease Morquio A. For the development of effective therapeutic approaches against this disease, the design of suitable enzyme enhancers (i.e. pharmacological chaperones) is fundamental. The natural substrates of GALNS are the glycosaminoglycans keratan sulfate and chondroitin 6-sulfate, which mainly display repeating units of sulfated carbohydrates. With a biomimetic approach, gold nanoparticles (AuNPs) decorated with simple monosaccharides, sulfated ligands (homoligand AuNPs), or both monosaccharides and sulfated ligands (mixed-ligand AuNPs) were designed here as multivalent inhibitors of GALNS. Among the homoligand AuNPs, the most effective inhibitors of GALNS activity are the β-D-galactoside-coated AuNPs. In the case of mixed-ligand AuNPs, β-D-galactosides/sulfated ligands do not show better inhibition than the β-D-galactoside-coated AuNPs. However, a synergistic effect is observed for α-D-mannosides in a mixed-ligand coating with sulfated ligands that reduced IC50 by one order of magnitude with respect to the homoligand α-D-mannoside-coated AuNPs. SAXS experiments corroborated the association of GALNS with β-D-galactoside AuNPs. These AuNPs are able to restore the enzyme activity by almost 2-fold after thermal denaturation, indicating a potential chaperoning activity towards GALNS. This information could be exploited for future development of nanomedicines for Morquio A. The recent implications of GALNS in cancer and neuropathic pain make these kinds of multivalent bionanomaterials of great interest towards multiple therapies.
2023-08-31 | Synthesis and Evaluation of Novel Triaryl Derivatives with Readthrough-Inducing Activity
The readthrough mechanism, which skips the premature termination codon and restores the biosynthesis of the defective enzyme, is an emerging therapeutic tactic for nonsense mutation-related diseases, such as Hurler syndrome, a type of mucopolysaccharidosis. In the present study, novel triaryl derivatives were synthesized and their readthrough-inducing activities were evaluated by a luciferase reporter assay with a partial α-L-iduronidase (IDUA) DNA sequence containing the Q70X nonsense mutation found in Hurler syndrome and by measuring the enzyme activity of IDUA knockout cells transfected with the mutant IDUA gene. KY-516, a representative compound in which the meta position carboxyl group of the left ring of the clinically used ataluren was converted to the para position sulfamoylamino group, the central ring to triazole, and the right ring to cyanobenzene, exhibited the most potent readthrough-inducing activity in the Q70X/luciferase reporter assay. In Q70X mutant IDUA transgenic cells, KY-516 significantly increased enzyme activity at 0.1 µM. After the oral administration of KY-516 (10 mg/kg), the highest plasma concentration of KY-516 was above 5 µM in rats. These results indicate that KY-516, a novel triaryl derivative, exhibits potent readthrough-inducing activity and has potential as a therapeutic agent for Hurler syndrome.
2020-12-11 | Molecular Insights into Determinants of Translational Readthrough and Implications for Nonsense Suppression Approaches
The fidelity of protein synthesis, a process shaped by several mechanisms involving specialized ribosome regions and external factors, ensures the precise reading of sense and stop codons. However, premature termination codons (PTCs) arising from mutations may, at low frequency, be misrecognized and result in PTC suppression, named ribosome readthrough, with production of full-length proteins through the insertion of a subset of amino acids. Since some drugs have been identified as readthrough inducers, this fidelity drawback has been explored as a therapeutic approach in several models of human diseases caused by nonsense mutations. Here, we focus on the mechanisms driving translation in normal and aberrant conditions, the potential fates of mRNA in the presence of a PTC, as well as on the results obtained in the research of efficient readthrough-inducing compounds. In particular, we describe the molecular determinants shaping the outcome of readthrough, namely the nucleotide and protein context, with the latter being pivotal to produce functional full-length proteins. Through the interpretation of experimental and mechanistic findings, mainly obtained in lysosomal and coagulation disorders, we also propose a scenario of potential readthrough-favorable features to achieve relevant rescue profiles, representing the main issue for the potential translatability of readthrough as a therapeutic strategy.
2020-07-31 | Screening, Synthesis, and Evaluation of Novel Isoflavone Derivatives as Inhibitors of Human Golgi β-Galactosidase
The genes GLB1 and GALC encode GLB1 isoform 1 and galactocerebrosidase, respectively, which exhibit β-galactosidase activity in human lysosomes. GLB1 isoform 1 has been reported to play roles in rare lysosomal storage diseases. Further, its β-galactosidase activity is the most widely used biomarker of senescent and aging cells; hence, it is called senescence-associated β-galactosidase. Galactocerebrosidase plays roles in Krabbe disease. We previously reported a novel β-galactosidase activity in the Golgi apparatus of human cells; however, the protein responsible for this activity could not be identified. Inhibitor-derived chemical probes can serve as powerful tools to identify the responsible protein. In this study, we first constructed a cell-based high-throughput screening (HTS) system for Golgi β-galactosidase inhibitors, and then screened inhibitors from two compound libraries using the HTS system, in vitro assay, and cytotoxicity assay. An isoflavone derivative was identified among the final Golgi β-galactosidase inhibitor compound hits. Molecular docking simulations were performed to redesign the isoflavone derivative into a more potent inhibitor, and six designed derivatives were then synthesized. One of the derivatives, ARM07, exhibited potent inhibitory activity against β-galactosidase, with an IC50 value of 14.8 µM and competitive inhibition with Ki value of 13.3 µM. Furthermore, the in vitro and cellular inhibitory activities of ARM07 exceeded those of deoxygalactonojirimycin. ARM07 may contribute to the development of affinity-based chemical probes to identify the protein responsible for the newly discovered Golgi β-galactosidase activity. The therapeutic relevance of ARM07 against lysosomal storage diseases and its effect on senescent cells should be evaluated further.
cell therapies
2022-12-25 | Modifying enzyme replacement therapy – A perspective
Abstract Several diseases are caused by the lack of functional proteins, including lysosomal storage diseases or haemophilia A and B. Patients suffering from one of these diseases are treated via enzyme replacement therapies to restore the missing protein. Although this treatment strategy prevents some disease symptoms, enzyme replacement therapies are very expensive and require very frequent infusions, which can cause infusion adverse reactions and massively impair the quality of life of the patients. This review proposes a technology to sustainably produce proteins within the patient to potentially make frequent protein‐infusions redundant. This technology is based on blood circulating immune cells as producers of the needed therapeutic protein. To ensure a stable protein concentration over time the cells are equipped with a system, which induces cell proliferation when low therapeutic protein levels are detected and a system inhibiting cell proliferation when high therapeutic protein levels are detected.
2019-06-18 | A decade of excellent transplant survival in children with inherited metabolic diseases: A report from a single metabolic transplant centre in Europe
Hematopoietic cell transplantation (HCT) confers a long-term disease-modifying therapy for transplant-permissive inherited metabolic diseases (IMDs). We examined the overall survival (OS) and engrafted survival (ES) of children with IMDs, who received first HCT at Royal Manchester Children's hospital from 1985 to 2016. A total of 137 children with IMDs were included in this analysis (historical cohort [1985-2006], n＝65; current cohort [2007-2016], n＝72). Primary diagnoses included mucopolysaccharidoses (81%), X-linked adrenoleukodystrophy (6%), metachromatic leukodystrophy (4%), mannosidosis (3%), Wolman disease (2%), and other conditions (4%). The five-year OS has increased from 65% (95% confidence interval [CI], 52%-76%) in the historical cohort to 91% (95% CI, 81%-96%) in the current cohort (P<0.001). Moreover, the five-year ES, which was 64% (95 CI%, 56%-72%) for the entire cohort, has doubled from 41% (95% CI, 29%-53%) in the historical cohort to 85% (95% CI, 75%-92%) in the current cohort (P<0.001). The proportion of patients with graft failure has decreased from 37% in the historical cohort to 8% in the current cohort (P<0.001). In patients who received a second transplant, 13 out of 20 patients (65%) in the historical cohort and all four in the current cohort were alive and engrafted. Of 82 survivors followed-up at Manchester, 80% and 20% had full and mixed chimerism, respectively. Although this study was restricted to a single center, our findings show that HCT is an increasingly safe procedure and provides long-lasting endogenous enzyme replacement therapy for children with IMDs in the modern era of HCT.
2019-02-01 | Preliminary results demonstrate engraftment with minimal neutropenia with MGTA-456, a CD34expanded cord blood (CB) product in patients transplanted for inherited metabolic disorders (IMD)
Mucopolysaccharidosis type IH (Hurler Syndrome), metachromatic leukodystrophy (MLD), globoid cell leukodystrophy (GLD) and cerebral adrenoleukodystrophy (cALD) are progressive diseases treatable with allogeneic hematopoietic stem cell transplantation (HSCT). While cord blood is a common stem cell source, it may be associated with prolonged neutropenia and graft failure. MGTA-456 is a cell therapy produced from a single CB unit using an aryl hydrocarbon receptor antagonist in a 15-day expansion culture of CD34+cells. In previous studies, patients with hematologic malignancies demonstrated a median 324-fold expansion of CD34+cells, and the time to neutrophil recovery was significantly reduced by a median of 9 days compared to historical controls. As higher CD34+doses have been correlated with improved engraftment and outcomes in inherited metabolic disease patients, we initiated a Phase 2, open-label trial to enroll up to 12 patients with a diagnosis of MPS1, cALD, MLD, and GLD utilizing CB units matched at ≥6 of 8 HLA loci using allele-based typing. The HSCT regimen consists of anti-thymocyte globulin (days -9 to -6), fludarabine (days -5 to -2) and busulfan (days -5 to -2), with cyclosporin and methylprednisolone as graft vs. host disease prophylaxis. In four patients treated thus far, a marked expansion of CD34+cells (median 482-fold) was observed. Two patients had no days of neutropenia and 2 patients had 1 and 4 days respectively, in contrast to a mean of 7.8 days in a historical Minnesota cohort (N=27) with identical conditioning. Donor myeloid chimerism (≥98%) was achieved by day +14 in all patients. One patient developed autoimmune cytopenia, a known complication unrelated to MSGT 456, resulting in death at day +143. Based on these promising data, MGTA-456 has potential to improve hematopoietic cell recovery and transplant-related outcomes in patients undergoing HSCT for patients with Inherited metabolic disorders.
2017-04-29 | Delivery of Cargo to Lysosomes Using GNeosomes
Liposomes have been used to improve the intracellular delivery of a variety of cargos. Encapsulation of cargos in liposomes leads to improved plasma half-lives and minimized degradation. Here, we present a method for improving the selective delivery of liposomes to the lysosomes using a guanidinylated neomycin (GNeo) transporter. The method for synthesizing GNeo-lipids, incorporating them into liposomes, and the enhanced lysosomal delivery of encapsulated cargo are presented. GNeo-liposomes, termed GNeosomes, are capable of delivering a fluorescent dye to the lysosomes of Chinese hamster ovary cells as shown using confocal microscopy. GNeosomes can also be used to deliver therapeutic quantities of lysosomal enzymes to fibroblasts isolated from patients with a lysosomal storage disorder.
2016-09-15 | Cell Therapy of Corneal Diseases
Mesenchymal stem cells isolated from connective tissues are pluripotent and differentiate into phenotypes of connective tissue cell lineages (osteoblasts, chondrocytes, and adipocytes) in vitro and in vivo. They have been used to treat mouse models of connective tissue disease such as lumican-null (Lum−/−) and mucopolysaccharidosis (Gusb) mice. Mesenchymal stem cells have unique immunosuppressive properties allowing evasion of host rejection; thus, they are valuable tools for cell therapy of congenital and acquired diseases involving immune dysfunction of multiple tissues including ocular surface tissues (cornea). We previously showed that human umbilical mesenchymal stem cells (UMSCs) modulated host immune responses, enabling them to survive xenograft transplantation. In vitro, UMSCs modulated inflammatory cells by inhibiting adhesion and invasion, and inducing cell death. UMSCs also regulated M1/M2 macrophage polarization and induced T-regulatory cell maturation from naive intraperitoneal cavity lavage cells. UMSCs exposed to inflammatory cells synthesized a rich extracellular glycocalyx composed of hyaluronan (HA) bound to the heavy chains (HCs) of inter-alpha-trypsin inhibitor (HC-HA), which contains tumor necrosis factor-α-stimulated gene 6 (TSG6) that catalyzes the transfer of HCs to HA, versican, and pentraxin-3. Our in vivo and in vitro results showed that the glycocalyx regulated inflammatory cells, allowing UMSCs to survive host immune rejection. Administration of antibodies against glycocalyx constituents or digestion with hyaluronidase and chondroitinase ABC abolished the UMSCs' ability to modulate immune responses. Treatment with anti-CD44 antibodies also diminished modulation of M2 macrophages by UMSCs, indicating that cell surface CD44 is required for correct UMSC glycocalyx assembly to modulate inflammatory cells.
gene therapies
2024-02-09 | REMOVER-PITCh: microhomology-assisted long-range gene replacement with highly multiplexed CRISPR-Cas9
A variety of CRISPR-Cas9-based gene editing technologies have been developed, including gene insertion and gene replacement, and applied to the study and treatment of diseases. While numerous studies have been conducted to improve the efficiency of gene insertion and to expand the system in various ways, there have been relatively few reports on gene replacement technology; therefore, further improvements are still needed in this context. Here, we developed the REMOVER-PITCh system to establish an efficient long-range gene replacement method and demonstrated its utility at two genomic loci in human cultured cells. REMOVER-PITCh depends on microhomology-assisted gene insertion technology called PITCh with highly multiplexed CRISPR-Cas9. First, we achieved gene replacement of about 20-kb GUSB locus using this system. Second, by applying the previously established knock-in-enhancing platform, the LoAD system, along with REMOVER-PITCh, we achieved the replacement of a longer gene region of about 200 kb at the ARSB locus. Our REMOVER-PITCh system will make it possible to remove and incorporate a variety of sequences from and into the genome, respectively, which will facilitate the generation of various disease and humanized models.
2023-05-27 | Etiology Understanding Is the Core of Skeletal Diagnosis
Background: The term idiopathic is a non-specific disease label, which fails account for proper etiological understanding. The frequent usage of the term idiopathic may indicate clinical incompetence. We describe a number of patients from our practice with a history of complex bone disorders who were falsely given the term idiopathic by other Institutes. The sole tool used by other Institutes to conclude osteoporosis as a diagnosis is the bone mineral density. Material and Methods: Two Austrian families and one unrelated adult female patient (total number of 12 subjects) have been referred to our department with referral letters confirming the diagnosis of idiopathic osteoporosis. All patients manifested a diverse constellation of skeletal deformities. In all these patients, regular courses of anti-resorptive treatment plus calcium and vitamin D supplements. These have been excessively prescribed. Results: Our diagnostic process which is based heavily on detailed clinical and radiological phenotypic characterizations, we refuted the diagnosis of idiopathic osteoporosis for all patients because of the clear-cut clinical features. For the first family, we suggested the diagnosis of a novel type of mucopolysaccharidosis (the genetic results confirmed the diagnosis of a novel subtype of mucopolysaccharidosis (through the identification of arylsulfatase K ARSK). For the second family which has been subjected to a lengthy useless investigation for almost more than a decade, we refuted the diagnosis of idiopathic osteoporosis and the whole exome sequencing was consistent with the phenotypic diagnosis Nasu-Hakola disease (heterozygous missense mutations in TYROBP gene). The other unrelated 42-year-old- female patient was correctly diagnosed with Nasu-Hakola disease. Conclusion: The objective of this study is to minimize and suppress the usage of the term idiopathic. It has been imperative to understand the sequence of pathological events that occurred in these families. The core for diagnosis is the etiological understanding, which stemmed from professional conscientiousness. Patients underwent misdiagnosis of idiopathic osteoporosis and were treated extensively with anti-resorptive agents, calcium supplements and vitamin D. Strikingly, these medications resulted in a combination of serious cranial and cerebral lesions because of massive pathological calcifications. Imaging analysis via conventional radiographs, MRI and CT scan showed massive calcifications of the cranial sutures, inter-clinoid ligaments and hyper-calcification of the skull base. Sadly speaking, early onset dementia emerged in connection with sclerosing leukoencephalopathy. We believe that the exogenous toxic effects of the huge amounts of bone supplements consumed by these patients enhanced the pre-existing metabolic bone and cerebral disorders. Our findings can ameliorate the etiological understanding of early onset dementia and other correlated cranial and skeletal lesions.
2019-03-01 | Enzymatic replacement therapy for lysosomal storage disorders: Drug evaluations review in Spain
In the European Union companies only need to demonstrate that the risk-benefit balance of the new drug is favourable to obtain the authorization to sell new drugs. Hence a comparison with available treatments, a cost-effectiveness analysis and the place in the therapy of the new drug are not required. Therefore, it is necessary to carry out these analyses in an additional study. In Spain there is no national government agency, which conducts a centralized evaluation and makes decisions on funding and are the Pharmacy and Therapeutics committees that take mandatory decisions for the entire region. To identify all drug assessments and health technology assessment reports of the enzymatic replacement treatments for the four LSD considered in this study, including Mucopolysaccharidosis (MPS), Gaucher, Fabry and Pompe disease at the national, regional and hospital level; and to summarize the efficacy in terms of outcome measures, adverse events, economic impact and final recommendations and indicate potential improvements. 19 reviews at the regional and hospital level and one therapeutic positioning report at national level for MPS were considered for this. In general, the drugs evaluated in LSD have proved to be effective in the most frequent clinical forms of disease. However, there are clinical phenotypes that still do not have available an effective treatment. It is expected that new therapies, such as intrathecal therapies or gene therapy could be evaluated in the different types of LSD demonstrating positive effects, especially in the previous phenotypes.
2019-02-03 | Donor-Free Gene Correction by Targeted Interhomolog Recombination
Spontaneous gene correction by interhomolog recombination (IHR) occasionally occurs to ameliorate genetic diseases of blood and skin 1–3 . Using an engineered endogenous gene as a reporter, we demonstrate that gene correction by IHR is normally infrequent (≤0.02%) but is stimulated by DSBs targeted by CRISPR/Cas9 to both homologous chromosomes; reaching frequencies of 0.5%. We further show that depletion of POLQ stimulates IHR frequencies 4-fold, to 2%, and promotes IHR in G2 phase, when recombination between replicated homologs can correct not only compound heterozygous but also autosomal dominant “gain-of-function” mutations, which present a special challenge for gene therapy. The strategies reported here will enable optimization of IHR for gene therapy in a variety of cell types. Advantages include the ability to correct gain-of-function mutations, no need for an exogenous donor, and the potential to limit damage to coding sequence by targeting IHR to introns.
2018-12-27 | Basic and Clinical Application of Adeno-Associated Virus–Mediated Genome Editing
Traditional gene therapy (gene replacement) has made a breakthrough in treating inherited diseases. Adeno-associated virus (AAV) has emerged as a highly promising vector with innate ability, boosting the development of gene replacement and gene targeting. With the recent advance of engineered nucleases that work efficiently in human cells, AAV mediated-genome editing with nucleases has raised hopes for in situ gene therapy of inherited and non-inherited diseases. Here, the applications of AAV-mediated genome editing are highlighted, and the prospect of AAV and nucleases that will render extension of such success in clinical gene therapy is discussed.
Access all drug discovery papers and probability of success in trials forecasts:
Access all drug discovery papers and probability of success in trials forecasts:
Drug Discovery Landscape
0 orphan drug designations.
0 orphan drug designations.
Let's accelerate rare disease drug discovery
Let's accelerate drug discovery
Get access to Explority AI’s forecasts to outperform average preclinical success rates. Whether you’re expanding your R&D pipeline, evaluating a partnership, or simply have a question — we’d love to hear from you.