Development of iPSC-derived iNKT cells to promote hematopoietic engraftment

摘要 造血干细胞移植(HSCT)是一种治疗多种恶性肿瘤的潜在疗法。 以及其他血液疾病，也为艾滋病毒等传染病的治疗带来了希望。 然而，造血干细胞移植的并发症(如移植物衰竭)的发病率和死亡率很高。 (这导致对致命性感染的高度易感性)和白血病复发。改进的新策略 因此，造血移植的结果将对临床产生重大影响。这个项目的重点是 人诱导多能干细胞体外培养不变自然杀伤T细胞的研究 作为一种辅助造血干细胞移植的细胞疗法，旨在促进成功的造血植入。在最近 已发表的初步研究表明，将同种异体人CD4iNKT细胞添加到人类 脐带血移植显著改善免疫缺陷患者的造血植入 老鼠。INKT细胞的植入促进活性是由于它们与脐带血的相互作用 单核细胞能产生几种强大的促造血因子，包括细胞因子 GM-CSF和IL-3(由iNKT细胞产生)，以及PGE2(由单核细胞响应来自 INKT细胞)。先前对小鼠模型的研究已经证实，细胞因子程序和功能特性 INKT细胞受两个关键转录因子PLZF和E4BP4控制。这些转录因子是如何 细胞因子对人iNKT细胞的影响尚不清楚，尤其是对iNKT细胞的影响。 GM-CSF和IL-3的产生以及与单核细胞的相互作用尚不清楚。该项目的目标1建议 建立缺乏或过表达PLZF或E4BP4的人IPSC来源的iNKT细胞。目标2将决定 这些转录因子对IPSC来源的iNKT细胞促进人类免疫功能的影响 造血植入和移植物抗淋巴瘤(GVL)活性。通过这些研究，我们将获得 对PLZF和E4BP4表达对iNKT功能特性的影响的高度新颖的见解，我们将 构建表达修饰转录因子的IPSC来源的iNKT细胞的可行性 旨在稳定特定的功能程序。这将为设计基于IPSC的iNKT奠定基础 转录调节的细胞，以促进造血植入，而不会对GVL产生不利影响。

Summary Hematopoietic stem cell transplantation (HSCT) is a potentially curative treatment for a variety of malignancies and other hematological disorders, and also holds promise for treatment of infectious diseases such as HIV. However, HSCT is limited by high risks of morbidity and mortality from complications such as graft failure (which leads to high susceptibility to lethal infections) and leukemic relapse. New strategies to improve outcomes of hematopoietic transplantation would thus have a significant clinical impact. This project focuses on developing invariant Natural Killer T (iNKT) cells derived from human induced pluripotent stem cells (iPSCs) as an adjunct HSCT cellular therapy designed to promote successful hematopoietic engraftment. In recently published preliminary studies, we showed that addition of allogeneic human CD4+ iNKT cells to human umbilical cord blood transplants led to dramatically improved hematopoietic engraftment in immunodeficient mice. The engraftment-promoting activity of iNKT cells was due to their interactions with cord blood monocytes that resulted in production of several potently pro-hematopoietic factors, including the cytokines GM-CSF and IL-3 (produced by iNKT cells), and PGE2 (produced by monocytes in response to signals from iNKT cells). Prior studies in murine models have established that cytokine programs and functional properties of iNKT cells are controlled by two key transcription factors, PLZF and E4BP4. How these transcription factors influence the cytokine profiles of human iNKT cells remains unclear, and specifically, their impact on iNKT cell production of GM-CSF and IL-3 and interactions with monocytes is not known. Aim 1 of this project proposes to generate human iPSC-derived iNKT cells lacking or over-expressing PLZF or E4BP4. Aim 2 will determine the impact of these transcription factors on the ability of iPSC-derived iNKT cells to promote human hematopoietic engraftment and on graft-versus-lymphoma (GVL) activity. Through these studies we will gain highly novel insight into the impact of PLZF and E4BP4 expression on iNKT functional properties, and we will establish the feasibility of generating iPSC-derived iNKT cells with modified transcription factor expression designed to stabilize specific functional programs. This will lay a foundation for designing iPSC-derived iNKT cells that are transcriptionally tuned to promote hematopoietic engraftment without adversely affecting GVL.