Strategies for prenatal stem cell transplantation

描述（由申请人提供）：宫内造血细胞移植（IUHCT）是一种有前景的治疗方法，可能适用于大量遗传性疾病。利用IUHCT的小鼠模型，我们已经建立了跨越完整MHC屏障的混合造血嵌合体，并了解了植入和相关供体特异性耐受的要求。我们已经确定了IUHCT后耐受的机制，并利用产前耐受促进出生后骨髓移植和免疫治疗，使部分供体嵌合体转化为完全供体嵌合体，跨越完整的MHC屏障，而无毒性。我们的长期目标是将这些成功的策略转化为临床应用，并开发新的安全的新方法来实现单独使用IUHCT的治疗性植入。本研究的具体目标是：1）明确决定IUHCT后同种异体移植成功或失败的适应性免疫反应的组成部分和机制。我们已经观察到使用BM和HSC的同种和同种异体IUHCT之间的植入频率存在明显差异。这些差异已经被我们目前的血管内注射大剂量细胞的技术所揭示。在这个目标中，我们将调查的假设，许多供体反应性细胞逃脱克隆缺失，嵌合体的维持是依赖于产生足够的外周调节反应之前，拒绝供体细胞。2)通过特异性靶向产前免疫治疗增强植入。我们将研究使用主要在造血细胞上表达的次要组织相容性免疫显性抗原作为特异性免疫治疗的靶点，以增强无GVHD胎儿的植入。3)探讨提高供体细胞归巢和植入的选择性策略。我们还确定了通过阻断二肽基肽酶CD26来改善子宫内移植后供体细胞归巢和植入的机制。本提案中的研究旨在开发和优化IUHCT的新方法，这些方法将安全地转移到大型动物和人类系统中。这些研究的最终目标是开发安全有效的策略，可以转化为子宫内治疗广泛的人类遗传性血液病，包括镰状细胞病和地中海贫血。

DESCRIPTION (provided by applicant): In utero hematopoietic cell transplantation (IUHCT) is a promising therapeutic approach potentially applicable to a large number of genetic diseases. Utilizing the murine model of IUHCT, we have established mixed hematopoietic chimerism across full MHC barriers and gained understanding into the requirements for engraftment and associated donor specific tolerance. We have determined the mechanism of tolerance after IUHCT, and utilized prenatal tolerance to facilitate postnatal bone marrow transplantation and immunotherapy with conversion of partial to complete donor chimerism across full MHC barriers without toxicity. Our long-term objective is the translation of these successful strategies to clinical application, and the development of novel and safe new approaches to achieve therapeutic engraftment by IUHCT alone. In this proposal our specific aims are: 1) To define the components and mechanism of the adaptive immune response that determine the success or failure of allogeneic engraftment after IUHCT. We have observed that there are clear differences in the frequency of engraftment between congenic and allogeneic IUHCT using BM and HSC. These differences have been unmasked by our current technique of intravascular injection of large doses of cells. In this aim we will investigate the hypothesis that many donor reactive cells escape clonal deletion, and that the maintenance of chimerism is dependent upon generation of an adequate peripheral regulatory response prior to rejection of donor cells. 2) To enhance engraftment by specifically targeted prenatal immunotherapy. We will investigate the use of minor histocompatibility immunodominant antigens expressed predominantly on hematopoietic cells as a target for specific immunotherapy to enhance engraftment in the fetus without GVHD. 3) To investigate selective strategies to improve homing and engraftment of donor cells. We have also identified a mechanism to improve homing and engraftment of donor cells after in utero transplantation by blockade of the dipeptidylpeptidase CD26. The studies in this proposal are designed to develop and optimize novel approaches to IUHCT that will be safely translatable to large animal and human systems. The ultimate goal of these studies is to develop safe and effective strategies that can be translated to the in utero treatment of a broad range of human genetic hematologic disorders including Sickle Cell Disease and Thalassemia.