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.

项目概要 体细胞基因治疗为治疗多种其他疾病提供了一种有前途的治疗方法 绝症或严重致残疾病。基因组编辑技术的最新发展包括 CRISPR（成簇规则间隔短回文重复序列）系统使执行 细胞中的精确基因工程。然而，CRISPR技术在人类患者中的临床应用已经 由于使用传统的递送方法体内疗效不足，一直具有挑战性。因此，它是 迫切需要开发一个能够将体外精确基因组编辑与 工程细胞在体内的有效应用。 皮肤的表皮祖细胞具有多种独特的优势，使其特别适合 离体基因治疗。人体皮肤是体内最大、最容易接近的器官，因此很容易隔离 皮肤表皮祖细胞并监测组织是否存在潜在的有害并发症。从解剖学上来说，皮肤 表皮通过基底膜与脉管系统分开，这防止了潜在的传播 体内转基因细胞，使潜在的治疗组织具有特异性和安全性。最后，潜力 皮肤基因治疗的适用性很广泛，因为有充分的证据表明，表达的蛋白质 在皮肤表皮细胞中可以穿过表皮/真皮屏障到达循环达到治疗效果 以系统的方式。此外，皮肤表皮细胞代谢酶的异位表达可以 将工程皮肤转变为“代谢池”，以纠正各种代谢紊乱。然而， 尽管具有潜在的临床重要性，基于表皮祖细胞的疗法（皮肤基因 由于缺乏合适的小鼠模型，该疗法受到了极大的阻碍。尽管老鼠或人类的皮肤 可以移植到免疫缺陷小鼠体内，但由于缺乏完整的免疫系统，无法进行检查 该疗法可能在体内引起的潜在结果和并发症。我们现在已经解决了 攻克技术难关，建立了独特的鼠对鼠皮肤移植模型，可稳定导入 将基因组编辑的表皮祖细胞转化为具有免疫能力的小鼠。在这个提案中，我们将利用 这个新平台并探索皮肤基因疗法治疗皮肤病的可行性和临床潜力 遗传性疾病，包括苯丙酮尿症 (PKU) 和 A 型血友病。我们的研究将共同​​建立一个 利用当前基因组编辑技术进行皮肤基因治疗的独特而强大的模型，揭示了 表皮祖细胞体细胞基因治疗的治疗潜力。