Pub Date : 2024-03-04DOI: 10.1016/j.omtm.2024.101229
Citra N.Z. Mattar, Wei Leong Chew, Poh San Lai
Gene modification therapies (GMTs) are slowly but steadily making progress toward clinical application. As the majority of rare diseases have an identified genetic cause, and as rare diseases collectively affect 5% of the global population, it is increasingly important to devise gene correction strategies to address the root causes of the most devastating of these diseases and to provide access to these novel therapies to the most affected populations. The main barriers to providing greater access to GMTs continue to be the prohibitive cost of developing these novel drugs at clinically relevant doses, subtherapeutic effects, and toxicity related to the specific agents or high doses required. strategy and treating younger patients at an earlier course of their disease could lower these barriers. Although currently regarded as niche specialties, prenatal and preconception GMTs offer a robust solution to some of these barriers. Indeed, treating either the fetus or embryo benefits from economy of scale, targeting pre-pathological tissues in the fetus prior to full pathogenesis, or increasing the likelihood of complete tissue targeting by correcting pluripotent embryonic cells. Here, we review advances in embryo and fetal GMTs and discuss requirements for clinical application.
{"title":"Embryo and fetal gene editing: Technical challenges and progress toward clinical applications","authors":"Citra N.Z. Mattar, Wei Leong Chew, Poh San Lai","doi":"10.1016/j.omtm.2024.101229","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101229","url":null,"abstract":"Gene modification therapies (GMTs) are slowly but steadily making progress toward clinical application. As the majority of rare diseases have an identified genetic cause, and as rare diseases collectively affect 5% of the global population, it is increasingly important to devise gene correction strategies to address the root causes of the most devastating of these diseases and to provide access to these novel therapies to the most affected populations. The main barriers to providing greater access to GMTs continue to be the prohibitive cost of developing these novel drugs at clinically relevant doses, subtherapeutic effects, and toxicity related to the specific agents or high doses required. strategy and treating younger patients at an earlier course of their disease could lower these barriers. Although currently regarded as niche specialties, prenatal and preconception GMTs offer a robust solution to some of these barriers. Indeed, treating either the fetus or embryo benefits from economy of scale, targeting pre-pathological tissues in the fetus prior to full pathogenesis, or increasing the likelihood of complete tissue targeting by correcting pluripotent embryonic cells. Here, we review advances in embryo and fetal GMTs and discuss requirements for clinical application.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"21 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140156561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Peptide drug discovery has great potential, but the cell membrane is a major obstacle when the target is an intracellular protein‒protein interaction (PPI). It is difficult to target PPIs with small molecules; indeed, there are no intervention tools that can target any intracellular PPI. In this study, we developed a platform that enables the introduction of peptides into cells via mRNA-based gene delivery. Peptide-length nucleic acids do not enable stable ribosome binding and exhibit little to no translation into protein. In this study, a construct was created in which the sequence encoding dihydrofolate reductase (DHFR) was placed in front of the sequence encoding the target peptide, together with a translation skipping sequence, as a sequence that meets the requirements of promoting ribosome binding and rapid decay of the translated protein. This enabled efficient translation from the mRNA encoding the target protein while preventing unnecessary protein residues. Using this construct, we showed that it can inhibit Drp1/Fis1 binding, one of the intracellular PPIs, which governs mitochondrial fission, an important aspect of mitochondrial dynamics. In addition, it was shown to inhibit pathological hyperfission, normalize mitochondrial dynamics and metabolism, and inhibit apoptosis of the mitochondrial pathway.
{"title":"Peptide-encoding gene transfer to modulate intracellular protein‒protein interactions","authors":"Toshihiko Taya, Daisuke Kami, Fumiya Teruyama, Satoaki Matoba, Satoshi gojo","doi":"10.1016/j.omtm.2024.101226","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101226","url":null,"abstract":"Peptide drug discovery has great potential, but the cell membrane is a major obstacle when the target is an intracellular protein‒protein interaction (PPI). It is difficult to target PPIs with small molecules; indeed, there are no intervention tools that can target any intracellular PPI. In this study, we developed a platform that enables the introduction of peptides into cells via mRNA-based gene delivery. Peptide-length nucleic acids do not enable stable ribosome binding and exhibit little to no translation into protein. In this study, a construct was created in which the sequence encoding dihydrofolate reductase (DHFR) was placed in front of the sequence encoding the target peptide, together with a translation skipping sequence, as a sequence that meets the requirements of promoting ribosome binding and rapid decay of the translated protein. This enabled efficient translation from the mRNA encoding the target protein while preventing unnecessary protein residues. Using this construct, we showed that it can inhibit Drp1/Fis1 binding, one of the intracellular PPIs, which governs mitochondrial fission, an important aspect of mitochondrial dynamics. In addition, it was shown to inhibit pathological hyperfission, normalize mitochondrial dynamics and metabolism, and inhibit apoptosis of the mitochondrial pathway.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"50 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140037415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-28DOI: 10.1016/j.omtm.2024.101227
Steven J. Hersch, Siddarth Chandrasekaran, Jamie Lam, Nafiseh Nafissi, Roderick A. Slavcev
Biotechnologies such as gene therapy have brought DNA vectors to the forefront of pharmaceuticals. The quality of starting material plays a pivotal role in determining final product quality. Here we examined the fidelity of DNA replication using enzymatic methods () compared to plasmid DNA produced in . Next-generation sequencing approaches rely on polymerases, which have inherent limitations in sensitivity. To address this challenge, we introduce a novel assay based on loss-of-function (LOF) mutations in the conditionally toxic gene. Our findings show that DNA production in results in significantly fewer LOF mutations (80- to 3000-fold less) compared to enzymatic DNA replication methods such as PCR and rolling circle amplification (RCA). These results suggest that using DNA produced by PCR or RCA may introduce a substantial number of mutation impurities, potentially affecting the quality and yield of final pharmaceutical products. Our study underscores that DNA synthesized has a significantly higher mutation rate than DNA produced traditionally in . Therefore, utilizing enzymatically-produced DNA in biotechnology and biomanufacturing may entail considerable fidelity-related risks, while using DNA starting material derived from substantially mitigates this risk.
基因治疗等生物技术将 DNA 载体推向了制药业的前沿。起始材料的质量在决定最终产品质量方面起着至关重要的作用。在这里，我们研究了使用酶法（）复制 DNA 的保真度，并与......中生产的质粒 DNA 进行了比较。下一代测序方法依赖于聚合酶，而聚合酶在灵敏度方面存在固有的局限性。为了应对这一挑战，我们引入了一种基于条件毒性基因功能缺失（LOF）突变的新型检测方法。我们的研究结果表明，与聚合酶链式反应（PCR）和滚动圈扩增（RCA）等酶DNA复制方法相比，DNA生产过程中产生的LOF突变明显较少（少80-3000倍）。这些结果表明，使用 PCR 或 RCA 生产的 DNA 可能会引入大量突变杂质，从而可能影响最终药品的质量和产量。我们的研究强调，合成 DNA 的突变率明显高于传统方法生产的 DNA。因此，在生物技术和生物制造中使用酶法生产的 DNA 可能会带来相当大的与保真度相关的风险，而使用来自的 DNA 起始材料则大大降低了这种风险。
{"title":"Manufacturing DNA in E. coli yields higher fidelity DNA than in vitro enzymatic synthesis","authors":"Steven J. Hersch, Siddarth Chandrasekaran, Jamie Lam, Nafiseh Nafissi, Roderick A. Slavcev","doi":"10.1016/j.omtm.2024.101227","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101227","url":null,"abstract":"Biotechnologies such as gene therapy have brought DNA vectors to the forefront of pharmaceuticals. The quality of starting material plays a pivotal role in determining final product quality. Here we examined the fidelity of DNA replication using enzymatic methods () compared to plasmid DNA produced in . Next-generation sequencing approaches rely on polymerases, which have inherent limitations in sensitivity. To address this challenge, we introduce a novel assay based on loss-of-function (LOF) mutations in the conditionally toxic gene. Our findings show that DNA production in results in significantly fewer LOF mutations (80- to 3000-fold less) compared to enzymatic DNA replication methods such as PCR and rolling circle amplification (RCA). These results suggest that using DNA produced by PCR or RCA may introduce a substantial number of mutation impurities, potentially affecting the quality and yield of final pharmaceutical products. Our study underscores that DNA synthesized has a significantly higher mutation rate than DNA produced traditionally in . Therefore, utilizing enzymatically-produced DNA in biotechnology and biomanufacturing may entail considerable fidelity-related risks, while using DNA starting material derived from substantially mitigates this risk.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"53 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140036798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-27DOI: 10.1016/j.omtm.2024.101225
Jonathan Al-Saadi, Mathias Waldén, Mikael Sandell, Jesper Solmér, Rikard Grankvist, Ida Friberger, Agneta Andersson, Mattias Carlsten, Kenneth Chien, Johan Lundberg, Nevin Witman, Staffan Holmin
Heart failure has a poor prognosis and no curative treatment exists. Clinical trials are investigating gene- and cell-based therapies to improve cardiac function. The safe and efficient delivery of these therapies to solid organs is challenging. Herein, we demonstrate the feasibility of using an endovascular intramyocardial delivery approach to safely administer mRNA drug products and perform cell transplantation procedures in swine. Using a -vessel wall (TW) device, we delivered chemically modified mRNAs (modRNA) and mRNA-enhanced mesenchymal stromal cells expressing vascular endothelial growth factor A (VEGF-A) directly to the heart. We monitored and mapped the cellular distribution, protein expression, and safety tolerability of such an approach. The delivery of modRNA-enhanced cells via the TW device with different flow rates and cell concentrations marginally affect cell viability and protein expression . Implanted cells were found within the myocardium for at least 3 days following administration, without the use of immunomodulation and minimal impact on tissue integrity. Finally, we could increase the protein expression of VEGF-A over 500-fold in the heart using a cell-mediated modRNA delivery system compared with modRNA delivered in saline solution. Ultimately, this method paves the way for future research to pioneer new treatments for cardiac disease.
{"title":"Endovascular transplantation of mRNA-enhanced mesenchymal stromal cells results in superior therapeutic protein expression in swine heart","authors":"Jonathan Al-Saadi, Mathias Waldén, Mikael Sandell, Jesper Solmér, Rikard Grankvist, Ida Friberger, Agneta Andersson, Mattias Carlsten, Kenneth Chien, Johan Lundberg, Nevin Witman, Staffan Holmin","doi":"10.1016/j.omtm.2024.101225","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101225","url":null,"abstract":"Heart failure has a poor prognosis and no curative treatment exists. Clinical trials are investigating gene- and cell-based therapies to improve cardiac function. The safe and efficient delivery of these therapies to solid organs is challenging. Herein, we demonstrate the feasibility of using an endovascular intramyocardial delivery approach to safely administer mRNA drug products and perform cell transplantation procedures in swine. Using a -vessel wall (TW) device, we delivered chemically modified mRNAs (modRNA) and mRNA-enhanced mesenchymal stromal cells expressing vascular endothelial growth factor A (VEGF-A) directly to the heart. We monitored and mapped the cellular distribution, protein expression, and safety tolerability of such an approach. The delivery of modRNA-enhanced cells via the TW device with different flow rates and cell concentrations marginally affect cell viability and protein expression . Implanted cells were found within the myocardium for at least 3 days following administration, without the use of immunomodulation and minimal impact on tissue integrity. Finally, we could increase the protein expression of VEGF-A over 500-fold in the heart using a cell-mediated modRNA delivery system compared with modRNA delivered in saline solution. Ultimately, this method paves the way for future research to pioneer new treatments for cardiac disease.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"90 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140097485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-27DOI: 10.1016/j.omtm.2024.101224
Isabella Elias Yonezawa Ogusuku, Vera Herbel, Simon Lennartz, Caroline Brandes, Eva Argiro, Caroline Fabian, Carola Hauck, Conny Hoogstraten, Sabrina Veld, Lois Hageman, Karin Teppert, Georgia Koutsoumpli, Marieke Griffioen, Nadine Mockel-Tenbrinck, Thomas Schaser, Rosa de Groot, Ian C.D. Johnston, Dominik Lock
Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and for subgroups with poor prognosis. A recently discovered T cell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1) presents an attractive option for the development of a cancer antigen-targeted cellular therapy. Manufacturing of TCR-modified T cells, however, is still limited by a complex, time-consuming, and laborious procedure. Therefore, this study specifically addressed the requirements for a scaled manufacture of ΔNPM1-specific T cells in an automated, closed, and good manufacturing practice-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive T cells were enriched, activated, lentivirally transduced, expanded, and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from 12 to 8 days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered T cells. The cellular product mainly consisted of highly viable CD8-positive T cells with an early memory phenotype. ΔNPM1-TCR CD8 T cells manufactured with the optimized process showed specific killing of AML and . The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.
急性髓性白血病（AML）是一种异质性恶性肿瘤，需要进一步改进治疗方法，尤其是针对老年人和预后不良的亚群。最近发现的一种靶向突变型核嗜磷蛋白 1（ΔNPM1）的 T 细胞受体（TCR）为开发癌症抗原靶向细胞疗法提供了一种极具吸引力的选择。然而，TCR修饰T细胞的制造仍然受到复杂、耗时和费力的程序的限制。因此，本研究特别针对ΔNPM1特异性T细胞的规模化生产要求，采用了自动化、封闭式和符合良好生产规范的流程。从低温保存的白细胞开始，对 2E8 CD8 阳性 T 细胞进行富集、激活、慢病毒转导、扩增，最后进行配制。通过调整和优化培养条件，我们还将制造时间从 12 天缩短到了 8 天，同时还获得了高达 5.5E9 ΔNPM1 TCR 工程 T 细胞的临床相关产量。细胞产品主要由具有早期记忆表型的高活性 CD8 阳性 T 细胞组成。用优化工艺制造的ΔNPM1-TCR CD8 T细胞对急性髓细胞白血病和白血病有特异性杀伤作用。该工艺已用于即将开展的治疗 NPM1 突变 AML 的 1/2 期临床试验。
{"title":"Automated manufacture of ΔNPM1 TCR-engineered T cells for AML therapy","authors":"Isabella Elias Yonezawa Ogusuku, Vera Herbel, Simon Lennartz, Caroline Brandes, Eva Argiro, Caroline Fabian, Carola Hauck, Conny Hoogstraten, Sabrina Veld, Lois Hageman, Karin Teppert, Georgia Koutsoumpli, Marieke Griffioen, Nadine Mockel-Tenbrinck, Thomas Schaser, Rosa de Groot, Ian C.D. Johnston, Dominik Lock","doi":"10.1016/j.omtm.2024.101224","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101224","url":null,"abstract":"Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and for subgroups with poor prognosis. A recently discovered T cell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1) presents an attractive option for the development of a cancer antigen-targeted cellular therapy. Manufacturing of TCR-modified T cells, however, is still limited by a complex, time-consuming, and laborious procedure. Therefore, this study specifically addressed the requirements for a scaled manufacture of ΔNPM1-specific T cells in an automated, closed, and good manufacturing practice-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive T cells were enriched, activated, lentivirally transduced, expanded, and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from 12 to 8 days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered T cells. The cellular product mainly consisted of highly viable CD8-positive T cells with an early memory phenotype. ΔNPM1-TCR CD8 T cells manufactured with the optimized process showed specific killing of AML and . The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"127 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140097488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1016/j.omtm.2024.101216
Jyoti Rana, Roland W. Herzog, Maite Muñoz-Melero, Kentaro Yamada, Sandeep RP. Kumar, Ahn K. Lam, David M. Markusic, Dongsheng Duan, Cox Terhorst, Barry J. Byrne, Manuela Corti, Moanaro Biswas
{"title":"B Cell Focused Transient Immune Suppression Protocol for Efficient AAV Readministration to the Liver","authors":"Jyoti Rana, Roland W. Herzog, Maite Muñoz-Melero, Kentaro Yamada, Sandeep RP. Kumar, Ahn K. Lam, David M. Markusic, Dongsheng Duan, Cox Terhorst, Barry J. Byrne, Manuela Corti, Moanaro Biswas","doi":"10.1016/j.omtm.2024.101216","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101216","url":null,"abstract":"","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"52 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139926264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1016/j.omtm.2024.101217
Manuela Braun, Claudia Lange, Philipp Schatz, Brian Long, Johannes Stanta, Boris Gorovits, Edit Tarcsa, Vibha Jawa, Tong-Yuan Yang, Wibke Lembke, Nicole Miller, Fraser McBlane, Louis Christodoulou, Daisy Yuill, Mark Milton
{"title":"Preexisting antibody assays for gene therapy: Considerations on clinical cut-off for patient selection and companion diagnostic requirements","authors":"Manuela Braun, Claudia Lange, Philipp Schatz, Brian Long, Johannes Stanta, Boris Gorovits, Edit Tarcsa, Vibha Jawa, Tong-Yuan Yang, Wibke Lembke, Nicole Miller, Fraser McBlane, Louis Christodoulou, Daisy Yuill, Mark Milton","doi":"10.1016/j.omtm.2024.101217","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101217","url":null,"abstract":"","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"261 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139926242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-17DOI: 10.1016/j.omtm.2024.101213
Paritha Arumugam, Brenna C. Carey, Kathryn A. Wikenheiser-Brokamp, Jeffrey Krischer, Matthew Wessendarp, Kenjiro Shima, Claudia Chalk, Jennifer Stock, Yan Ma, Diane Black, Michelle Imbrogno, Margaret Collins, Dan Justin Kalenda Yombo, Haripriya Sakthivel, Takuji Suzuki, Carolyn Lutzko, Jose A. Cancelas, Michelle Adams, Elizabeth Hoskins, Dawn Lowe-Daniels, Lilith Reeves, Anne Kaiser, Bruce C. Trapnell
Pulmonary macrophage transplantation (PMT) is a gene and cell transplantation approach in development as therapy for hereditary pulmonary alveolar proteinosis (hPAP), a surfactant accumulation disorder caused by mutations in (and murine homologs). We conducted a toxicology study of PMT of gene-corrected macrophages (mGM-RαMϕs) or saline-control intervention in or wild-type (WT) mice including single ascending dose and repeat ascending dose studies evaluating safety, tolerability, pharmacokinetics, and pharmacodynamics. Lentiviral-mediated cDNA transfer restored GM-CSF signaling in mGM-RαMϕs. Following PMT, mGM-RαMϕs engrafted, remained within the lungs, and did not undergo uncontrolled proliferation or result in bronchospasm, pulmonary function abnormalities, pulmonary or systemic inflammation, anti-transgene product antibodies, or pulmonary fibrosis. Aggressive male fighting caused a similarly low rate of serious adverse events in saline- and PMT-treated mice. Transient, minor pulmonary neutrophilia and exacerbation of pre-existing hPAP-related lymphocytosis were observed 14 days after PMT of the safety margin dose but not the target dose (5,000,000 or 500,000 mGM-RαMϕs, respectively) and only in mice but not in WT mice. PMT reduced lung disease severity in mice. Results indicate PMT of mGM-RαMϕs was safe, well tolerated, and therapeutically efficacious in mice, and established a no adverse effect level and 10-fold safety margin.
{"title":"A toxicology study of Csf2ra complementation and pulmonary macrophage transplantation therapy of hereditary PAP in mice","authors":"Paritha Arumugam, Brenna C. Carey, Kathryn A. Wikenheiser-Brokamp, Jeffrey Krischer, Matthew Wessendarp, Kenjiro Shima, Claudia Chalk, Jennifer Stock, Yan Ma, Diane Black, Michelle Imbrogno, Margaret Collins, Dan Justin Kalenda Yombo, Haripriya Sakthivel, Takuji Suzuki, Carolyn Lutzko, Jose A. Cancelas, Michelle Adams, Elizabeth Hoskins, Dawn Lowe-Daniels, Lilith Reeves, Anne Kaiser, Bruce C. Trapnell","doi":"10.1016/j.omtm.2024.101213","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101213","url":null,"abstract":"Pulmonary macrophage transplantation (PMT) is a gene and cell transplantation approach in development as therapy for hereditary pulmonary alveolar proteinosis (hPAP), a surfactant accumulation disorder caused by mutations in (and murine homologs). We conducted a toxicology study of PMT of gene-corrected macrophages (mGM-RαMϕs) or saline-control intervention in or wild-type (WT) mice including single ascending dose and repeat ascending dose studies evaluating safety, tolerability, pharmacokinetics, and pharmacodynamics. Lentiviral-mediated cDNA transfer restored GM-CSF signaling in mGM-RαMϕs. Following PMT, mGM-RαMϕs engrafted, remained within the lungs, and did not undergo uncontrolled proliferation or result in bronchospasm, pulmonary function abnormalities, pulmonary or systemic inflammation, anti-transgene product antibodies, or pulmonary fibrosis. Aggressive male fighting caused a similarly low rate of serious adverse events in saline- and PMT-treated mice. Transient, minor pulmonary neutrophilia and exacerbation of pre-existing hPAP-related lymphocytosis were observed 14 days after PMT of the safety margin dose but not the target dose (5,000,000 or 500,000 mGM-RαMϕs, respectively) and only in mice but not in WT mice. PMT reduced lung disease severity in mice. Results indicate PMT of mGM-RαMϕs was safe, well tolerated, and therapeutically efficacious in mice, and established a no adverse effect level and 10-fold safety margin.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"27 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140569488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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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