Mechanisms whereby IFN-gamma sensitizes AML to the graft-vs-leukemia effect

IFN-γ 使 AML 对移植物抗白血病效应敏感的机制

• 批准号: 10609407
• 负责人: Warren D Shlomchik
• 金额: $ 60.46万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-20 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10609407
• 关键词: Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Address; Affinity; Alloantigen; Allogenic; Allografting; Biological Models; Biology; Blast Phase; Bone Marrow Cells; CD8B1 gene; CRISPR/Cas technology; CSPG6 gene; Cause of Death; Cells; Chronic-Phase Myeloid Leukemia; Clinic; Clinical; Clinical Data; Clustered Regularly Interspaced Short Palindromic Repeats; Cytomegalovirus; DNMT3a; Data; Donor person; FLT3 gene; Failure; Future; Gene Deletion; Gene Expression; Genes; Genetic; HOXA9 gene; Hematologic Neoplasms; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Heterogeneity; Human; IFNGR1 gene; Immune; Intercellular adhesion molecule 1; Interferon Type II; Interferons; KRAS2 gene; Knock-in; Knock-out; Knockout Mice; Knowledge; Leukemic Cell; MLL-AF9; Mediating; Minor Histocompatibility Antigens; Modeling; Molecular Abnormality; Mus; NUP98 gene; Natural Killer Cells; Neoplasms; Oncogenes; Patients; Persons; Phenocopy; Property; Recurrent disease; Relapse; Resistance; Role; STAT1 gene; STAT2 gene; Specimen; Stem cell transplant; T cell response; T cell therapy; T-Lymphocyte; TP53 gene; Testing; Transplantation; Work; acute myeloid leukemia cell; cell killing; cellular transduction; clinical application; curative treatments; experimental study; graft vs host disease; graft vs leukemia effect; leukemia; leukemia relapse; minor H antigen H60; molecular subtypes; mouse model; overexpression; pleiotropism; post-transplant; single-cell RNA sequencing; stem cells; therapeutic target; tool

Abstract. Allogeneic hematopoietic stem cell transplantation (alloSCT) is a curative therapy most commonly applied in treatment of patients with acute myeloblastic leukemia (AML). The progeny of alloreactive ab T cells in the allograft can kill recipient leukemia cells, thereby mediating the graft-vs-leukemia effect (GVL). Nevertheless, disease relapse, indicative of insufficient GVL, is the single most common cause of death post-transplant. GVL-resistance and sensitivity are not equal across classes of leukemias, despite all expressing alloantigens. For example, chronic phase chronic myelogenous leukemia (CP-CML) is exquisitely GVL-sensitive whereas blast crisis CML (BC-CML) is GVL-resistant, despite sharing common biology. Likewise, AML and acute lymphoblastic leukemia are relatively GVL-resistant. Because GVL-resistance and -sensitivity track with the class of neoplasm, we reasoned that they are leukemia cell-intrinsic properties. We therefore established GVL against mouse models of mCP-CML and mBC-CML created with authentic human oncogenes. Through the creation of gene-deficient leukemias we discovered that for effective GVL, mBC-CML requires IFN-γR stimulation whereas GVL is fully intact against STAT1/STAT2-/- mCP-CML that cannot respond to any type of IFN. IFN-γR-/- MLL-AF9 was also GVL-resistant. These data indicate that effective GVL against myeloblastic leukemias requires a high magnitude alloreactive T cell response that generates IFN-g whereas a more smoldering T cell response that does not create much IFN-g can be effective against CP-CML. These results also strongly suggest that the delivery of IFN-g, in conjunction with alloreactive or leukemia-reactive T cells, would have a major clinical impact. However, key questions need to be answered to optimally apply IFN-g or agents that induce some of its effects in the clinic. First, while all AML subtypes share some common biology, there is substantial heterogeneity in driver genes. It will therefore be critical to understand which AML molecular subtypes require IFN-g for optimal GVL. We aim to do so by using CRISPR-Cas9 to delete the IFN- γR from transplantable leukemias that develop in compound gene-edited mice that express key AML-driver genes, sometimes in conjunction with a gene deletion. We have already successfully edited the IFN-γR from JAK2V617F/p53-/- and FLT3-ITD/DNMT3-/-/NPM1c AMLs. Second, IFN-g is a blunt tool with pleiotropic effects that could promote graft-vs-host disease. Therefore, we aim to understand under what alloimmune conditions IFN-g is required and what specific downstream effects are essential to sensitize AML cells to GVL. We have already developed approaches to restore expression of key IFN-g-induced genes (e.g. CIITA, NLRC5, ICAM-1) to IFN-γR-/- leukemias in order to specifically interrogate their relevance. Further targets will be discovered through single cell RNAseq, focusing on IFN-g-induced gene expression changes in subpopulations with stem cell qualities. These targets could then be deleted or expressed in IFN-γR leukemias to further define their importance.

抽象的。 同种异体造血干细胞移植（同种）是一种治疗疗法，最常用于 治疗急性粒细胞白血病（AML）的患者。同种反应性AB T细胞的后代 同种异体移植可以杀死受体白血病细胞，从而介导移植物vs-白血病效应（GVL）。尽管如此， 疾病的缓解表明GVL不足，是移植后死亡最常见的原因。 在白血病类别中，GVL的抗性和敏感性并不相等，所有表达同型抗原的人。 例如，慢性期慢性粒细胞性白血病（CP-CML）恰恰是GVL敏感的 爆炸危机CML（BC-CML）具有GVL耐药性，Dospite共享共同的生物学。同样，AML和急性 淋巴细胞白血病相对抗GVL。因为GVL耐GVL和 - 敏感性轨迹 肿瘤类别，我们认为它们是白血病细胞中性特性。因此，我们建立了GVL 针对由正宗人类癌基因产生的MCP-CML和MBC-CML的小鼠模型。通过 基因缺陷的白血病的创建我们发现，对于有效的GVL，MBC-CML需要IFN-γR 刺激，而GVL完全完整地针对STAT1/STAT2 - / - MCP-CML，无法响应任何类型的 ifn。 IFN-γR - / - MLL-AF9也具有GVL耐药性。这些数据表明有效的GVL针对粒细胞 白血病需要高度同种反应性T细胞响应，该反应产生IFN-G，而更多 闷烧的T细胞响应不会产生太多IFN-G可以有效地针对CP-CML。这些结果 还强烈建议，IFN-G的递送以及同种异体反应性或白血病反应性T细胞的递送， 将产生重大的临床影响。但是，需要回答关键问题，以最佳地应用IFN-G或 影响其在诊所中某些影响的药物。首先，虽然所有AML亚型具有一些共同的生物学，但 驱动基因存在很大的异质性。因此，要了解哪个AML至关重要 分子亚型需要IFN-G才能获得最佳GVL。我们的目标是通过使用CRISPR-CAS9删除IFN- 来自可移植白血病的γR，在表达关键AML驱动器的复合基因编辑的小鼠中发育 基因有时与基因缺失结合。我们已经成功编辑了IFN-γR JAK2V617F/P53 - / - 和FLT3-ITD/DNMT3 - / - /NPM1C AMLS。其次，IFN-G是具有多效效应的钝工具 这可能会促进移植-VS宿主疾病。因此，我们旨在了解在什么同种免疫条件下 需要IFN-G，以及哪些特定的下游效应对于将AML细胞感知到GVL至关重要。我们有 已经开发了恢复关键IFN-G诱导基因表达的方法（例如CIITA，NLRC5，ICAM-1） 为了特别询问其相关性，以ifn-γr - / - 白血病。将发现进一步的目标 通过单细胞RNASEQ，重点介绍IFN-G诱导的基因表达在茎的亚群中的变化 细胞质量。然后可以在IFN-γR白血病中删除或表达这些靶标，以进一步定义其 重要性。