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

基本信息

批准号：
10217590
负责人：
Warren D Shlomchik
金额：
$ 58.51万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
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/10217590
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 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 Lead 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

项目摘要

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。异基因造血干细胞移植（alloSCT）是一种最常用于以下疾病的治疗方法：治疗急性髓细胞性白血病 (AML) 患者的同种异体反应性 ab T 细胞的后代。同种异体移植物可以杀死受体白血病细胞，从而介导移植物抗白血病效应（GVL）。疾病复发（GVL 不足的指标）是移植后最常见的死亡原因。尽管所有白血病都表达同种异体抗原，但不同类型的白血病的 GVL 抗性和敏感性并不相同。例如，慢性期慢性粒细胞白血病 (CP-CML) 对 GVL 极其敏感，而尽管急变性慢性粒细胞白血病 (BC-CML) 与急性髓性白血病 (AML) 具有相同的生物学特性，但它具有 GVL 抗性。淋巴细胞白血病对GVL 具有相对抵抗性，因为GVL 抵抗性和敏感性与GVL 相关。类肿瘤，我们推断它们是白血病细胞固有的特性，因此我们建立了 GVL。针对使用真实人类癌基因创建的 mCP-CML 和 mBC-CML 小鼠模型。基因缺陷型白血病的产生我们发现，为了有效的 GVL，mBC-CML 需要 IFN-γR 刺激，而 GVL 对 STAT1/STAT2-/- mCP-CML 完全完整，不能对任何类型的刺激做出反应 IFN-γR-/- MLL-AF9 也具有 GVL 抗性，这些数据表明 GVL 能有效对抗成髓细胞。白血病需要高强度的同种异体反应性 T 细胞反应，产生 IFN-g，而更多的不会产生大量 IFN-g 的闷烧 T 细胞反应可以有效对抗 CP-CML。还强烈建议将 IFN-g 与同种异体反应性或白血病反应性 T 细胞结合使用，然而，需要回答关键问题才能最佳地应用 IFN-g 或。首先，虽然所有 AML 亚型都有一些共同的生物学特性，因此，了解哪些 AML 具有显着的异质性。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-g 来实现这一目标。来自可移植白血病的 γR，在表达关键 AML 驱动因素的复合基因编辑小鼠中发育基因，有时与基因删除一起我们已经成功地编辑了 IFN-γR。 JAK2V617F/p53-/- 和 FLT3-ITD/DNMT3-/-/NPM1c AML 其次，IFN-g 是一种具有多效性的钝性工具。因此，我们的目标是了解在什么同种免疫条件下。 IFN-g 是必需的，并且哪些特定的下游效应对于使 AML 细胞对 GVL 敏感至关重要。已经开发出恢复关键 IFN-g 诱导基因表达的方法（例如 CIITA、NLRC5、ICAM-1） IFN-γR-/- 白血病，以具体询问其相关性，并将发现更多靶点。通过单细胞RNAseq，重点关注干细胞亚群中IFN-g诱导的基因表达变化然后可以在 IFN-γR 白血病中删除或表达这些靶标，以进一步定义它们。重要性。