Targeting a Novel Hematopoietic Stem Cell Population in Non-Human Primates for Effective and Sustained Gene Therapy

针对非人类灵长类动物的新型造血干细胞群进行有效和持续的基因治疗

基本信息

批准号：
10593456
负责人：
HANS-PETER KIEM
金额：
$ 68.47万
依托单位：
FRED HUTCHINSON CANCER CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-11-10 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10593456
关键词：
Affect Allogenic Animals Autologous Autologous Transplantation Blood Blood Cells Blood Platelets Bone Marrow CCR5 gene CD34 gene CRISPR/Cas technology Cells Clinical Clonality Data Disease Engraftment Family Flow Cytometry Frequencies Gene-Modified Genes Genetic Diseases Genetic Transcription Goals Grant Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hemoglobinopathies Human Immune Immune system Immunologic Deficiency Syndromes In Vitro Lentivirus Life Maintenance Mediating Outcome Patients Phenotype Population Property Recovery Regimen Safety Siblings Source Stem cell transplant Supplementation Technology Testing Time Translating Transplantation Umbilical Cord Blood Whole-Body Irradiation base conditioning cost curative treatments gene therapy graft vs host disease hematopoietic stem cell expansion high throughput screening improved in vivo neutrophil nonhuman primate novel peripheral blood side effect small molecule stem cell gene therapy stem cell population stem cells success transcriptome sequencing transplant model treatment strategy zinc finger nuclease zinc finger nuclease technology

项目摘要

Project Summary Autologous hematopoietic stem cell (HSC) gene therapy is one of the most promising strategies for the treatment of genetic diseases affecting the blood and immune system. Currently, the only curative treatment for patients with diseases like hemoglobinopathies or immunodeficiencies is an allogeneic HSC transplant. While outcomes with an HLA-matched sibling donor are very encouraging, most patients will not have an HLA-matched family donor. HSC transplants from unrelated donors or partially matched donors can be associated with life- threatening side effects especially from graft-vs-host disease (GVHD) and related infectious complications. To avoid these complications, efforts have been made to use the patients´ own (autologous) HSCs and correct them with either lentivirus-mediated gene therapy or gene editing. A critical problem and bottleneck, however, has been the inability to identify and purify a “true” HSC population and therefore optimize conditions for such cells that allow for rapid multi-lineage engraftment. A reliable strategy to determine the quality and quantity of such a HSC population would be a major advance for high-throughput screening of ex vivo gene editing and expansion conditions. Using our nonhuman primate (NHP) stem cell transplantation model, we have recently identified an exclusive HSC-enriched population capable of multi-lineage short-term and long-term engraftment that is evolutionary conserved between human and NHPs. This HSC-enriched population accounts for ~3-5% of the entire CD34+ cell population, reducing the number of cells for gene therapy approaches by ~25-fold. Most importantly, flow-cytometric quantification of this HSC-enriched phenotype allowed us to reliably predict engraftment success as well as the onset of neutrophil and platelet recovery after HSC transplantation regardless of the source, gene modification, or ex vivo expansion. Availability of this novel HSC-enriched phenotype in combination with the ability to easily enrich for this subpopulation will help overcome current limitations of autologous HSC gene therapy. We hypothesize that gene editing of this HSC-enriched population will improve targeting, enhance gene editing efficiency, and increase in vivo persistence of gene-modified and corrected blood and immune cells. Furthermore, we hypothesize that this phenotype will serve as a reliable read-out to develop ex vivo culture conditions for the expansion of gene-modified HSCs promoting improved engraftment, which in turn should enable us to reduce the intensity of currently used conditioning regimens for HSC gene therapy. Evolutionary conservation of this HSC-enriched phenotype between human and NHP cells will allow us to rapidly translate these findings to clinical HSC gene therapy studies.

项目概要自体造血干细胞（HSC）基因治疗是最有前途的治疗策略之一目前，治疗影响血液和免疫系统的遗传性疾病是患者的唯一治疗方法。血红蛋白病或免疫缺陷等疾病是同种异体 HSC 移植的结果。与 HLA 匹配的兄弟姐妹捐赠者的情况非常令人鼓舞，大多数患者不会有 HLA 匹配的家庭来自无关捐献者或部分匹配捐献者的 HSC 移植可能与生命有关。威胁性副作用，尤其是移植物抗宿主病（GVHD）和相关感染并发症。为了避免这些并发症，已努力使用患者自己的（自体）HSC 并纠正它们然而，无论是慢病毒介导的基因治疗还是基因编辑，都存在一个关键问题和瓶颈。无法识别和纯化“真正的”HSC 群体，因此无法优化此类细胞的条件允许快速多谱系植入的可靠策略来确定这种质量和数量。 HSC群体将成为离体基因编辑和扩增高通量筛选的重大进步使用我们的非人类灵长类动物 (NHP) 干细胞移植模型，我们最近发现了一种。富含 HSC 的群体能够进行独特的多谱系短期和长期移植，即人类和 NHP 之间的进化保守性，这种富含 HSC 的群体约占总数的 3-5%。整个 CD34+ 细胞群，将基因治疗方法的细胞数量减少了约 25 倍。重要的是，这种富含 HSC 的表型的流式细胞术定量使我们能够可靠地预测移植成功以及 HSC 移植后中性粒细胞和血小板恢复的开始，无论这种富含 HSC 的新表型的来源、基因修饰或离体扩增的可用性。与轻松丰富该亚群的能力相结合将有助于克服目前的局限性我们追求的是，对富含 HSC 的人群进行基因编辑将得到改善。靶向性，提高基因编辑效率，增加基因修饰和校正的体内持久性此外，我们追求这种表型将作为可靠的读数。开发用于扩增基因修饰 HSC 的体外培养条件，促进改善植入，这反过来应该使我们能够减少目前使用的 HSC 基因调理方案的强度人类和 NHP 细胞之间这种富含 HSC 的表型的进化保守性将使我们能够进行治疗。快速将这些发现转化为临床 HSC 基因治疗研究。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Safe and efficient lentiviral vector integration with HSC-targeted gene therapy.

DOI：
10.1182/bloodadvances.2022009087
发表时间：
2023-09-12
期刊：
BLOOD ADVANCES
影响因子：
7.5
作者：
Radtke, Stefan;Pande, Dnyanada;Enstrom, Mark;Kiem, Hans-Peter
通讯作者：
Kiem, Hans-Peter

AMD3100 redosing fails to repeatedly mobilize hematopoietic stem cells in the nonhuman primate and humanized mouse.

DOI：
10.1016/j.exphem.2020.11.001
发表时间：
2021-01
期刊：
Experimental hematology
影响因子：
2.6
作者：
Samuelson C;Radtke S;Cui M;Perez A;Kiem HP;Humbert O
通讯作者：
Humbert O

Efficient long-term multilineage engraftment of CD33-edited hematopoietic stem/progenitor cells in nonhuman primates.

DOI：
10.1016/j.omtm.2023.101121
发表时间：
2023-12-14
期刊：
MOLECULAR THERAPY METHODS & CLINICAL DEVELOPMENT
影响因子：
0
作者：
Petty, Nicholas E.;Radtke, Stefan;Fields, Emily;Humbert, Olivier;Llewellyn, Mallory J.;Laszlo, George S.;Zhu, Haiying;Jerome, Keith R.;Walter, Roland B.;Kiem, Hans-Peter
通讯作者：
Kiem, Hans-Peter

Stochastic fate decisions of HSCs after transplantation: early contribution, symmetric expansion, and pool formation.

移植后 HSC 的随机命运决定：早期贡献、对称扩展和池形成。

DOI：
10.1182/blood.2022018564
发表时间：
2023
期刊：
Blood
影响因子：
20.3
作者：
Radtke,Stefan;Enstrom,Mark;Pande,Dnyanada;Duke,ElizabethR;Cardozo-Ojeda,EFabian;Madhu,Ravishankar;Owen,Staci;Kanestrom,Greta;Cui,Margaret;Perez,AnaiM;Schiffer,JoshuaT;Kiem,Hans-Peter
通讯作者：
Kiem,Hans-Peter

Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells.

非人灵长类造血干细胞和祖细胞的鉴定。