Development of epidermal progenitor cell-based therapy for regenerative medicine

开发基于表皮祖细胞的再生医学疗法

• 批准号: 9280088
• 负责人: Xiaoyang Wu
• 金额: $ 38.39万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9280088
• 关键词: Anatomy; Animal Model; Animals; Antibodies; Autologous Transplantation; Basement membrane; Blood Circulation; Blood Coagulation Disorders; Blood Coagulation Factor; Blood coagulation; Cell Therapy; Cells; Classical phenylketonuria; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cutaneous; Dermal; Development; Disease; Ectopic Expression; Engineering; Engraftment; Enzymes; Epidermis; Epilepsy; Factor VIII; Future; Gene Delivery; Genetic Engineering; Hemophilia A; Hepatic; Hereditary Disease; Human; Immune system; Immunocompetent; Immunodeficient Mouse; In Vitro; Inborn Genetic Diseases; Inherited; Mediating; Metabolic; Metabolic Diseases; Modeling; Monitor; Mus; Mutation; Organ; Organoids; Outcome; Patients; Phenotype; Phenylalanine; Phenylalanine Ammonia-Lyase; Phenylalanine Hydroxylase; Phenylketonurias; Plasma; Problem behavior; Procedures; Process; Production; Proteins; Psyche structure; Regenerative Medicine; Research; Safety; Skin; Skin Transplantation; Somatic Gene Therapy; Specificity; Stem cell transplant; Stem cells; System; Technology; Testing; Therapeutic; Therapeutic Effect; Tissue Therapy; Tissues; Transplantation; Treatment Efficacy; Work; adult stem cell; amino acid metabolism; clinical application; clinical development; clinically relevant; disability; disabling disease; efficacy testing; gene therapy; genetically modified cells; genome editing; human disease; immunoreaction; in vivo; keratinocyte; mouse model; novel; precise genome editing; prevent; regenerative therapy; spatiotemporal; stem cell therapy

Project Summary Somatic gene therapy provides a promising therapeutic approach for treatment of a variety of otherwise terminal or severely disabling diseases. The recent development of genome editing technology, including CRISPR (clustered regularly-interspaced short palindromic repeats) system, has made it possible to perform precise genetic engineering in cells. However, clinical application of CRISPR technology to human patients has been challenging due to the inadequate efficacy in vivo using conventional delivery approach. Thus, it is urgently needed to develop an ex vivo platform that can combine both precise genome editing in vitro with effective application of engineered cells in vivo. The epidermal progenitor cells of skin have several unique advantages, making it particularly suited for ex vivo gene therapy. Human skin is the largest and most accessible organ in the body, making it easy to isolate skin epidermal progenitor cells and monitor the tissue for potential detrimental complications. Anatomically, skin epidermis is separated from vasculature by the basement membrane, which prevents potential dissemination of genetically modified cell in vivo, making the potential therapy tissue specific and safe. Lastly, the potential applicability of cutaneous gene therapy is broad because it has been well documented that proteins expressed in skin epidermal cells can cross the epidermal/dermal barrier and reach circulation to achieve therapeutic effect in a systematic manner. In addition, ectopic expression of metabolic enzymes in skin epidermal cells can transform the engineered skin into a “metabolic sink” for correction of various metabolic disorders. However, despite the potential clinical importance, research in epidermal progenitor cell-based therapy (cutaneous gene therapy) has been greatly hindered due to lack of an appropriate mouse model. Although mouse or human skin can be transplanted to immunodeficient mice, lack of an intact immune system makes it impossible to examine the potential outcomes and complications that the therapy may elicit in vivo. We have now resolved the technical hurdle and established a unique mouse-to-mouse skin transplantation model that can stably introduce genome-edited epidermal progenitor cells into immunocompetent mice. In this proposal, we will take advantage of this novel platform and explore the feasibility and clinical potential of cutaneous gene therapy for treatment of genetic diseases, including phenylketonuria (PKU) and hemophilia A. Together, our studies will establish a unique and powerful model for cutaneous gene therapy with current genome editing technology, revealing the therapeutic potential for somatic gene therapy with epidermal progenitor cells.

项目摘要 体细胞基因疗法提供了一种有希望的治疗方法来治疗多种换 终端或严重致残的疾病。基因组编辑技术的最新发展，包括 CRISPR（聚集经常间隔的短圆柱式重复序列）系统使得能够执行 细胞中的精确基因工程。但是，CRISPR技术在人类患者中的临床应用已有 由于使用常规交付方法在体内效率不足，我们受到了挑战。那是 迫切需要开发一个可以在体外精确基因组编辑结合的离体平台 在体内有效应用工程细胞。 皮肤的表皮祖细胞具有几个独特的优势，使其特别适合 离体基因疗法。人体皮肤是体内最大，最容易接近的器官，使其易于分离 皮肤表皮祖细胞并监测组织是否有害并发症。解剖学，皮肤 表皮与基底膜与脉管系统分离，这阻止了潜在的传播 在体内转基因细胞，使潜在的治疗组织特异性且安全。最后，潜力 皮肤基因疗法的适用性很广泛，因为它已经有充分的证明是表达的蛋白质 在皮肤中，表皮细胞可以越过表皮/皮肤屏障并达到循环以实现热效应 以系统的方式。另外，皮肤表皮细胞中代谢酶的依托型表达可以 将工程的皮肤转变为“代谢水槽”，以纠正各种代谢疾病。然而， 尽管具有潜在的临床重要性，但表皮祖细胞细胞疗法的研究（皮肤基因 由于缺乏合适的小鼠模型，疗法受到了极大的阻碍。虽然老鼠或人类皮肤 可以移植到免疫缺陷的小鼠，缺乏完整的免疫系统，因此无法检查 治疗可能在体内引起的潜在结果和并发症。我们现在解决了 技术障碍并建立了一个独特的鼠标到鼠标皮肤移植模型，可以稳定引入 基因组编辑的表皮祖细胞进入免疫能力小鼠。在此提案中，我们将利用 在这个新型平台上，探索皮肤基因治疗治疗的可行性和临床潜力 遗传疾病，包括苯基酮尿（PKU）和血友病A。一起，我们的研究将建立一个 使用当前基因组编辑技术的皮肤基因治疗的独特而有力的模型，揭示了 表皮祖细胞体细胞基因治疗的治疗潜力。