Innovative Cell Therapy for Pediatric Acute Myeloid Leukemia

摘要 母R21提案1 R21 CA 245468 - 01 A1“儿科急性髓系白血病的创新细胞疗法” 旨在阐明儿童急性髓系白血病（pAML）母细胞对LV-10耐药的机制 细胞介导的杀伤LV-10细胞是工程化的1型调节性T细胞（Tr 1），代表了一种有前途的治疗方法 因为它们固有的抗髓系肿瘤细胞毒性，但在初步实验中， pAML母细胞抵抗LV-10细胞杀伤。RNA-seq揭示了两个可能导致pAML的候选基因 抵抗杀戮。这些基因也可以用作耐药pAML的生物标志物，这对于治疗急性白血病非常重要。 LV-10细胞的下游临床翻译。然而，需要更大的RNA-seq数据集来充分地识别RNA。 动力生物标志物发现。母提案的目标1将评估45例新pAML原始细胞对 LV-10介导的杀伤，除了中试样品之外，还为生物标志物发现提供了足够的能力。 补充提案的目的1将对这45个pAML原始细胞进行批量RNAseq分析，以确定 潜在的新的耐药性生物标志物。此外，LV-10细胞目前使用健康的细胞的混合物扩增。 人供体来源的外周血单核细胞（PBMC）作为饲养细胞，其将供体- LV-10扩增、表型和功能的供体变异性。在LV-10细胞的临床转化之前， 必须确定一种替代的人工扩张系统，该系统提供功能统一的GMP组件， 柔顺细胞产物。这些系统已经用于其他基于T细胞的疗法。在目标2的柔软- 我们将比较两种人工抗原呈递细胞（aAPC）系统，TransAct和 ImmunoCult，对传统的PBMC饲养细胞，LV-10细胞扩增，表型，细胞因子产生，去甲肾上腺素， 急性髓细胞白血病（AML）总之，这些数据将揭示：1）在pAML抗性中起作用的其他基因 LV-10杀伤，这可以在未来用于逆转这种耐药性或筛选符合条件的pAML患者。 LV-10细胞疗法;和2）用于LV-10细胞扩增的最佳aAPC系统。因此，本补充建议 将补充母提案的目标，它的重点是pAML转录组分析和一种新的细胞 治疗pAML。此外，执行实现这些目标所需的实验将提供 候选人在癌症免疫学和免疫治疗方面具有必要的背景和技能，为以下目标铺平道路 他的研究生入学和职业道路作为一个癌症免疫学家。

Abstract The parent R21 proposal 1R21CA245468-01A1 "Innovative Cell Therapy for Pediatric Acute Myeloid Leukemia" is designed to address the mechanism of pediatric acute myeloid leukemia (pAML) blast resistance to LV-10 cell-mediated killing. LV-10 cells are engineered type 1 regulatory T cells (Tr1) that represent a promising therapy for pAML because of their inherent anti-myeloid tumor cytotoxicity, but in pilot experiments, a subset of primary pAML blasts resisted LV-10 cell killing. RNA-seq revealed two candidate genes that could contribute to pAML resistance to killing. These genes could also be used as biomarkers of resistant pAML, which is important for downstream clinical translation of LV-10 cells. However, a larger RNA-seq dataset is required for sufficiently powered biomarker discovery. Aim 1 of the parent proposal will assess the sensitivity of 45 new pAML blasts to LV-10 mediated killing, which in addition to the pilot samples, provides sufficient power for biomarker discovery. Aim 1 of the supplemental proposal will perform bulk RNAseq analyses on these 45 pAML blasts to identify potential new biomarkers of resistance. Further, the LV-10 cells are currently expanded using a mixture of healthy human donor-derived peripheral blood mononuclear cells (PBMC) as feeder cells, which introduces donor-to- donor variability to LV-10 expansion, phenotype and function. Before the clinical translation of LV-10 cells, it is essential to identify an alternative artificial expansion system that provides a functionally uniform and GMP com- pliant cell product. These systems are already in place for other T cell-based therapies. In aim 2 of the supple- mental proposal, we will compare the effect of 2 artificial antigen-presenting cell (aAPC) systems, TransAct and ImmunoCult, to the traditional PBMC feeders, on LV-10 cell expansion, phenotype, cytokine production, degran- ulation and AML killing. Altogether, this data will reveal: 1) additional genes that play a role in pAML resistance to LV-10 killing, which can be used in the future to reverse this resistance or screen pAML patients eligible for LV-10 cell therapy; and 2) optimal aAPC system for LV-10 cell expansion. As such, this supplemental proposal will complement the aims of the parent proposal, and it's focus on pAML transcriptome analysis and a novel cell therapy for pAML. Furthermore , execution of the experiments required to achieve these goals will provide the Candidate with essential background and skills in cancer immunology and immunotherapy, paving the way to his enrollment into graduate school and a career path as a cancer immunologist.