Molecular TherapyPub Date : 2025-05-28DOI: 10.1016/j.ymthe.2025.05.022
Jerusha Naidoo, Yin Ren, Matthew Rocco, Eric Bielefeld, Allison O'Brien, Hsuan-Chih Kuo, Nathan McFarland, Matthew Avenarius, Manpreet Takhar, Geneva Frank, Victor Van Laar, Krystof Bankiewicz
{"title":"Delivery of AAV vectors to the superior olivary complex enables efficient adult cochlear transduction via axonal transport.","authors":"Jerusha Naidoo, Yin Ren, Matthew Rocco, Eric Bielefeld, Allison O'Brien, Hsuan-Chih Kuo, Nathan McFarland, Matthew Avenarius, Manpreet Takhar, Geneva Frank, Victor Van Laar, Krystof Bankiewicz","doi":"10.1016/j.ymthe.2025.05.022","DOIUrl":"10.1016/j.ymthe.2025.05.022","url":null,"abstract":"<p><p>Significant breakthroughs have been made in translation of adeno-associated virus (AAV) vectors for hearing disorders targeting auditory hair cells (HCs). In addition to HCs, spiral ganglion neurons (SGNs) are also impacted in a large number of sensorineural hearing loss cases in adults. However when administered directly into the cochlea in rodents aged older than P1-P3, AAV-mediated SGN transduction efficiency decreases dramatically. An efficient gene-delivery method to transduce adult SGNs is needed. Our group has a track record of utilizing axonal transport to transduce brain structures distal from the site of AAV injection. We investigated whether SGNs could be transduced in adult rats following intraparenchymal AAV administration to the olivary complex. Cochlear transduction was observed with the following common AAV serotypes expressing green fluorescent protein: AAV6, AAV9, AAV-Anc80, and AAV-PhP.B. Cochlear transduction was observed with all serotypes, but the cellular tropism and efficiency of gene transfer varied across the cochlear spiral (apex, middle, base) with different AAV serotypes, with some transducing both SGNs and HCs, while others transduced SGNs or HCs exclusively. This study provides proof of concept that AAV delivery to the olivary complex can efficiently deliver transgenes to SGNs in the adult mammalian cochlea.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Airway-applied mRNA vaccine needs tailored sequence design and high standard purification that removes devastating dsRNA contaminant.","authors":"Jingjing Zhang, Chao Li, Yuheng Liu, Rui Liao, Dian He, Lifeng Xu, Tingting Chen, Qin Xiao, Mingxing Luo, Yang Chen, Yali Li, Huaxing Zhu, Joseph Rosenecker, Xiaoyan Ding, Shuchen Pei, Shan Guan","doi":"10.1016/j.ymthe.2025.05.024","DOIUrl":"10.1016/j.ymthe.2025.05.024","url":null,"abstract":"<p><p>The development of mucosal mRNA vaccines is promising but extremely challenging. Major efforts have been focused on optimizing delivery systems, but it is still unknown whether the intrinsic quality of mRNA components significantly impacts the potency of airway-inoculated mRNA vaccines. Here, we systematically demonstrate that mucosal mRNA vaccine requires higher standards of purification and a tailor-designed sequence to fulfill its potency compared to its parenteral-route-inoculated counterpart. Double-stranded RNA (dsRNA) contaminants are prone to trigger the innate immune response in the airway that activates the mRNA degradation mechanism, thereby diminishing mRNA expression and subsequent antigen-specific immune responses. To address these challenges, we developed a strategy that combines optimized untranslated regions (UTRs) screened from endogenous genes of pulmonary cells with affinity chromatography-based purification, which effectively removed dsRNA contaminants. The optimized mRNA administered via the airway route not only demonstrated superior protein expression (30-fold increase) and reduced inflammation in the lung but also promoted robust adaptive immunity comprising significantly elevated systemic, cellular, and mucosal immune responses. This was in stark contrast to the intramuscular-injected counterpart that displayed less-pronounced benefits. Our findings offer new insights into the development of mucosal mRNA therapeutics from an overlooked but crucial perspective of optimizing mRNA components.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144183291","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-28DOI: 10.1016/j.ymthe.2025.05.030
Dan Lu, Hui-Yi Chu, Soo Park, Mark Landon, Masanao Tsuda, Earl Avramis, Carissa Dege, Thomas Dailey, Yijia Pan, Sandeep Kothapally Hanok, Matthew Denholtz, Ramzey Abujarour, Tom Lee, John Goulding, Martin Hosking, Jodie Goodridge, Eigen Peralta, Bahram Valamehr
{"title":"A novel CD3ε fusion receptor allows use of T-cell engagers on TCR-less allogeneic CAR T cells to improve activity and prevent antigen escape.","authors":"Dan Lu, Hui-Yi Chu, Soo Park, Mark Landon, Masanao Tsuda, Earl Avramis, Carissa Dege, Thomas Dailey, Yijia Pan, Sandeep Kothapally Hanok, Matthew Denholtz, Ramzey Abujarour, Tom Lee, John Goulding, Martin Hosking, Jodie Goodridge, Eigen Peralta, Bahram Valamehr","doi":"10.1016/j.ymthe.2025.05.030","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.05.030","url":null,"abstract":"<p><p>Chimeric antigen receptor (CAR) T-cell therapies have shown clinical success in treating hematologic malignancies. However, heterogenous target antigen expression can impair the durability of response. Combining CAR and T-cell engagers (TCEs) targeting additional tumor antigens can address tumor heterogeneity and antigen escape. In allogeneic settings, eliminating the T-cell receptor (TCR) of the adoptive T-cell therapy prevents graft versus host disease. However, the absence of TCR leads to loss of surface CD3 expression, preventing cooperative activity with CD3-directed TCEs. We utilized induced pluripotent stem cells (iPSCs) to support the required multiplexed editing, establish a renewable starting material for off-the-shelf manufacture, and create the desired TCR-less CAR<sup>+</sup> CD3<sup>+</sup> T cells. Here we illustrate surface expression of a CD3ε fusion receptor (CD3FR) in iPSC-derived CAR T (CAR iT) cells, enabling TCE-mediated targeting of diverse antigens. In vitro and in vivo, CD3FR<sup>+</sup> CAR iT cells demonstrated potent cytotoxic response and cooperative activity against mixed tumor lines and multiple antigens. CD3FR<sup>+</sup> iT cells were further engineered to secrete TCEs, eliminating the need for extra supplementation with TCEs. Collectively, the data highlights the ability to integrate TCEs with allogeneic CAR iT cells for multi-antigen targeting, overcoming tumor relapse, and supporting off-the-shelf therapy for patient access.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-27DOI: 10.1016/j.ymthe.2025.05.023
Ying Xu, Jun Li, Zihao Wang, Rongrong Lu, Yingying Liu, Min Wang, Hao Li, Rui Zhao, Weijun Feng
{"title":"Ablation of dysmorphic neurons is a safe and effective treatment for focal cortical dysplasia II.","authors":"Ying Xu, Jun Li, Zihao Wang, Rongrong Lu, Yingying Liu, Min Wang, Hao Li, Rui Zhao, Weijun Feng","doi":"10.1016/j.ymthe.2025.05.023","DOIUrl":"10.1016/j.ymthe.2025.05.023","url":null,"abstract":"<p><p>Focal cortical dysplasia type II (FCDII) is a leading cause of refractory epilepsy in children, yet treatment options remain limited. The most frequent genetic cause of FCDII is mosaic and somatic variants in genes of the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) pathway, leading to hyperactivation of mTOR signaling. The presence of dysmorphic neurons (DNs) resulting from hyperactive mTOR signaling is critical for the development of epilepsy in FCDII. One critical therapeutic challenge and opportunity for FCDII is to selectively eliminate DNs. Here, we developed two strategies to specifically ablate DNs in FCDII mouse models, and the results demonstrate that DN ablation is sufficient to both prevent and eliminate epilepsy in mice. Moreover, the associated neurobehavioral abnormalities were also reversed following treatment. Therefore, our study provides proof-of-concept evidence that DN ablation is a highly promising approach for curing FCDII in the future.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-27DOI: 10.1016/j.ymthe.2025.05.027
Kathy Steece-Collier, Margaret E Caulfield, Molly J Vander Werp, Scott J Muller, Jennifer A Stancati, Yaping Chu, Ivette M Sandoval, Timothy J Collier, Jeffrey H Kordower, Fredric P Manfredsson
{"title":"Disease-modifying, multidimensional efficacy of putaminal Ca<sub>V</sub>1.3-shRNA gene therapy in aged parkinsonism male and female macaques.","authors":"Kathy Steece-Collier, Margaret E Caulfield, Molly J Vander Werp, Scott J Muller, Jennifer A Stancati, Yaping Chu, Ivette M Sandoval, Timothy J Collier, Jeffrey H Kordower, Fredric P Manfredsson","doi":"10.1016/j.ymthe.2025.05.027","DOIUrl":"10.1016/j.ymthe.2025.05.027","url":null,"abstract":"<p><p>There remain several unmet clinical needs in Parkinson's disease (PD) including waning and incomplete efficacy of symptomatic therapies, development of medication side effects (i.e., levodopa-induced dyskinesias [LID]) and unfettered disease progression. Ca<sub>V</sub>1.3 calcium channels are therapeutic targets of intense interest in PD. We developed an RNA interference (RNAi)-based vector approach utilizing adeno-associated virus (AAV) expressing a short-hairpin (sh)RNA against Ca<sub>V</sub>1.3 channels to provide potent, target-specific silencing of these channels that become dysfunctional in the parkinsonian striatum. We report here unprecedented evidence that magnetic resonance imaging-guided intraputaminal AAV-Ca<sub>V</sub>1.3-shRNA in aged (25-29 years) male and female nonhuman primates with long-standing (8 months) advanced parkinsonian motor deficits results in a significant progressive reversal of functional deficits in the absence of pharmacotherapy, with some aspects including postural instability and motivation-based fine-motor performance returning to normal/pre-parkinsonian baseline. This contrasts maintenance of stable moderate-to-severe disability in those receiving the control/scrambled vector (AAV-SCR-shRNA). AAV-Ca<sub>V</sub>1.3-shRNA recipients also demonstrate maintained levodopa motor benefit lost in these aged, parkinsonian subjects receiving the AAV-SCR-shRNA vector, similar to end-stage PD. Last, AAV-Ca<sub>V</sub>1.3-shRNA recipients showed unprecedented, near-complete prevention of LID induction despite long-term (5.5 months), twice-daily, dose-escalation levodopa. The realization of these first-in-class multimodal gene therapy attributes in the clinic would represent a major therapeutic advancement for PD.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-27DOI: 10.1016/j.ymthe.2025.05.028
Mark Anthony B Casel, Jae-Woo Ahn, Hyunjoon Kim, Isaac Choi, Seung-Gyu Jang, Rare Rollon, Ho-Young Ji, Mina Yu, Seong Cheol Min, Min-Suk Song, Young Ki Choi
{"title":"Therapeutic applications of interface-mimicking peptides for targeting the SARS-CoV-2 NSP12-NSP8 RdRp complex.","authors":"Mark Anthony B Casel, Jae-Woo Ahn, Hyunjoon Kim, Isaac Choi, Seung-Gyu Jang, Rare Rollon, Ho-Young Ji, Mina Yu, Seong Cheol Min, Min-Suk Song, Young Ki Choi","doi":"10.1016/j.ymthe.2025.05.028","DOIUrl":"10.1016/j.ymthe.2025.05.028","url":null,"abstract":"<p><p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication depends on the NSP12-NSP8-NSP7 complex, which plays a critical role in enhancing RNA-dependent RNA polymerase (RdRp) activity. NSP8 is particularly essential, stabilizing the RdRp complex and supporting viral replication across diverse variants. To disrupt this crucial interaction, we designed four NSP8-derived peptides-N8-Pepα, N8-Pepα_cyc, N8-Pepβ, and N8-PepβD-targeting a key hotspot region within the NSP12-NSP8 interface that governs complex stability and processivity. In vitro assays demonstrated that these peptides effectively inhibit RdRp activity by disrupting the NSP12-NSP8 interaction, leading to significant reductions in SARS-CoV-2 replication in Vero E6 cells. Notably, intranasal administration of N8-Pepα or N8-Pepα_cyc (25 mg/kg) in Balb/c mice provided robust antiviral protection, alleviating weight loss and reducing mortality following challenge with a mouse-adapted SARS-CoV-2 strain. Both prophylactic and therapeutic treatments significantly lowered viral titers and minimized pathological damage in the nasal turbinates and lungs. These results highlight the NSP12-NSP8 interface as a novel and highly conserved target for antiviral therapy and establish NSP8-derived peptides, particularly N8-Pepα and N8-Pepα_cyc, as promising candidates for inhibiting RdRp complex formation and controlling SARS-CoV-2 replication.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144160421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-27DOI: 10.1016/j.ymthe.2025.05.025
Anna Maria Giudice, Sydney L Roth, Stephanie Matlaga, Evan Cresswell-Clay, Pamela Mishra, Patrick M Schürch, Kwame Attah M Boateng-Antwi, Minu Samanta, Guillem Pascual-Pasto, Vincent Zecchino, Timothy T Spear, Brendan McIntyre, Neil C Chada, Tingting Wang, Lingling Liu, Ruoning Wang, John T Wilson, Adam J Wolpaw, Kristopher R Bosse
{"title":"Reprogramming the neuroblastoma tumor immune microenvironment to enhance GPC2 CAR T cells.","authors":"Anna Maria Giudice, Sydney L Roth, Stephanie Matlaga, Evan Cresswell-Clay, Pamela Mishra, Patrick M Schürch, Kwame Attah M Boateng-Antwi, Minu Samanta, Guillem Pascual-Pasto, Vincent Zecchino, Timothy T Spear, Brendan McIntyre, Neil C Chada, Tingting Wang, Lingling Liu, Ruoning Wang, John T Wilson, Adam J Wolpaw, Kristopher R Bosse","doi":"10.1016/j.ymthe.2025.05.025","DOIUrl":"10.1016/j.ymthe.2025.05.025","url":null,"abstract":"<p><p>Poor tumor trafficking and the immunosuppressive tumor microenvironment (TME) limit chimeric antigen receptor (CAR) T cell efficacy in solid tumors, such as neuroblastoma. We previously optimized GPC2 CARs in human neuroblastoma xenografts leading to clinical translation; however, there have not been preclinical studies using immunocompetent models. Thus, here we generated murine GPC2 CAR T cells using the D3-GPC2-targeting single-chain variable fragment being utilized clinically (NCT05650749) and tested them in neuroblastoma syngeneic allografts. Immune-profiling of GPC2 CAR T cell-treated tumors revealed significant reprogramming of the TME, most notably poor intra-tumor CAR T cell persistence being associated with increased recruitment of myeloid-derived suppressor cells (MDSCs), along with MDSC-recruiting CXCL1/2 chemokines. These tumor-infiltrating MDSCs directly inhibited GPC2 CAR T cell activation, proliferation, and cytotoxicity ex vivo. To both capitalize on this chemokine gradient and mitigate MDSC-tumor trafficking, we engineered GPC2 CAR T cells to express the CXCL1/2 receptor, CXCR2. CXCR2-armored GPC2 CAR T cells migrated toward CXCL1/2 gradients, enhanced anti-neuroblastoma efficacy, and reduced the level of MDSCs in the TME. Together, these findings suggest CAR T cell studies in immunocompetent models are imperative to define mechanisms of solid tumor immune escape and rationally design armoring strategies that will lead to durable clinical efficacy.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-27DOI: 10.1016/j.ymthe.2025.05.026
Bin Li, Xudong Zhu, Jingyu Ge, Hangyu Zhang, Yang Gao, Xuanwen Bao, Xiaomeng Dai, Zhicheng Du, Xiaoxuan Tu, Zhou Tong, Qihan Fu, Dongxue Hu, Weihong Tian, Yi Zheng, Lulu Liu, Peng Zhao, Weijia Fang, Shu Wang, Dongrui Wang
{"title":"Intraperitoneal infusion of NKG2D CAR-NK cells induces endogenous CD8<sup>+</sup> T cell activation in patients with advanced colorectal cancer.","authors":"Bin Li, Xudong Zhu, Jingyu Ge, Hangyu Zhang, Yang Gao, Xuanwen Bao, Xiaomeng Dai, Zhicheng Du, Xiaoxuan Tu, Zhou Tong, Qihan Fu, Dongxue Hu, Weihong Tian, Yi Zheng, Lulu Liu, Peng Zhao, Weijia Fang, Shu Wang, Dongrui Wang","doi":"10.1016/j.ymthe.2025.05.026","DOIUrl":"10.1016/j.ymthe.2025.05.026","url":null,"abstract":"<p><p>Peritoneal metastasis (PM) is a prevalent mode of metastasis in colorectal cancer (CRC) with rapid disease progression and limited treatment options. Locoregional infusion of immune cells have been explored in different types of solid tumors, but the feasibility and safety of locoregional chimeric antigen receptor-natural killer (CAR-NK) cells in treating CRC-PM is still unknown. We report the results of a phase 1 dose-escalation clinical trial (NCT05213195) using intraperitoneal infusion of NKG2D CAR-NK cells in heavily-pretreated patients with CRC-PM, and the immune-modulatory effect of CAR-NK cells was also illustrated using multi-omics analysis and functional validation. Locoregional NKG2D CAR-NK cell therapy showed preliminary efficacy and tolerable toxicity in this study. Of nine patients, three achieved stable disease, and the disease control rate was 33.3%. Notably, CD8<sup>+</sup> T cells post CAR-NK infusion showed the enrichment of a subset co-expressing CD38 and human leukocyte antigen (HLA)-DR, which were highly clonal and also associated with a cytotoxic phenotype. This activated CD38<sup>+</sup> HLA-DR<sup>+</sup> T cell subset, together with their effector phenotype and function, was induced and maintained by the secretome of activated CAR-NK cells. Our data provide evidence supporting the feasibility of locoregionally infused CAR-NK cells to treat patients with CRC-PM, and indicate peripheral immune activation after locoregional CAR-NK therapy.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174194","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-24DOI: 10.1016/j.ymthe.2025.05.018
Francesca Del Bufalo,Concetta Quintarelli,Franco Locatelli
{"title":"Allogeneic CAR T cells: A new player in the field and the peculiar opportunities of the hospital exemption path.","authors":"Francesca Del Bufalo,Concetta Quintarelli,Franco Locatelli","doi":"10.1016/j.ymthe.2025.05.018","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.05.018","url":null,"abstract":"","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":"33 1","pages":""},"PeriodicalIF":12.4,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144136870","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular TherapyPub Date : 2025-05-21DOI: 10.1016/j.ymthe.2025.05.020
Raghavi Sudharsan,Leonardo Murgiano,Aditi Ahuja,Yu Sato,Jennifer Kwok,Natalia Dolgova,Svetlana Savina,Morgan Sedorovitz,Valerie L Dufour,Gustavo D Aguirre,Leah C Byrne,William A Beltran
{"title":"Novel Photoreceptor-Specific Promoters for Gene Therapy in Mid-to-Late Stage Retinal Degeneration.","authors":"Raghavi Sudharsan,Leonardo Murgiano,Aditi Ahuja,Yu Sato,Jennifer Kwok,Natalia Dolgova,Svetlana Savina,Morgan Sedorovitz,Valerie L Dufour,Gustavo D Aguirre,Leah C Byrne,William A Beltran","doi":"10.1016/j.ymthe.2025.05.020","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.05.020","url":null,"abstract":"Inherited retinal degenerations (IRDs) cause progressive photoreceptor loss, leading to vision impairment. Gene therapy using adeno-associated viral (AAV) vectors holds immense promise for treating these conditions. However, achieving optimal gene expression at mid-to-late stages of retinal degeneration remains challenging due to scarcity of efficient photoreceptor-specific promoters expressed at these disease stages. This study aimed to identify and validate novel promoters capable of robust and specific transgene expression when ≥ 50% photoreceptors are lost. Analysis of transcriptomic data from two naturally occurring canine IRD models, laser capture microdissection of retinal cryosections followed by qPCR, and RNA in situ hybridization identified six promising genes with sustained or upregulated expression in photoreceptors in late-stage disease. Upstream cis-regulatory elements of both canine and human orthologs were identified and characterized using in silico analyses and dual-luciferase assays. Short promoters (≤ 840 bp) derived from GNGT2, IMPG2 and PDE6H genes exhibited robust reporter gene expression in photoreceptors when delivered via AAV to the subretinal space of two non-allelic canine IRD models at mid- and late- disease stages. These findings provide a strategy to enhance AAV-mediated gene therapy by enabling sustained transgene expression in degenerating retinas, improving treatment outcomes for patients with progressive vision loss.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":"57 1","pages":""},"PeriodicalIF":12.4,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144122066","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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