MPN-inducing mutations as biomarkers of synthetic lethality

MPN 诱导突变作为合成致死率的生物标志物

• 批准号: 10174883
• 负责人: TOMASZ SKORSKI
• 金额: $ 42.77万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10174883
• 关键词: Acute Myelocytic Leukemia; Affect; BRCA deficient; BRCA mutations; Back; Base Excision Repairs; Biological Markers; Blood; Cell Survival; Cells; Chronic Phase; DNA; DNA Double Strand Break; DNA Repair; DNA Repair Disorder; DNA Repair Gene; DNA Single Strand Break; DNA replication fork; DNA-PKcs; DNA-dependent protein kinase; Defect; Disease; Disease remission; Double Strand Break Repair; Down-Regulation; Drug Targeting; EZH2 gene; Epigenetic Process; FDA approved; Foundations; Gene Mutation; Genes; Genetic; Hemorrhagic Thrombocythemia; Immunodeficient Mouse; Induced Mutation; JAK1 gene; JAK2 gene; Knock-in; Knock-out; LIG4 gene; MPL gene; Malignant - descriptor; Malignant Neoplasms; Mediating; Modality; Modeling; Mus; Mutate; Mutation; Mutation Analysis; Myeloproliferative disease; Myelosuppressive Therapy; New Agents; Nonhomologous DNA End Joining; Normal Cell; Normal tissue morphology; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Polycythemia Vera; Primary Myelofibrosis; Prognostic Marker; Proliferating; Published Comment; Reactive Oxygen Species; Reporting; Research; Role; Testing; Therapeutic; Therapeutic Effect; Time; Toxic effect; Update; Xenograft procedure; base; brca gene; calreticulin; cancer cell; cancer therapy; driver mutation; homologous recombination; humanized mouse; hydroxyurea; improved; in vivo evaluation; individual patient; inhibitor/antagonist; kinase inhibitor; leukemia/lymphoma; mouse model; novel therapeutic intervention; novel therapeutics; potential biomarker; pre-clinical; prevent; recombinational repair; repaired; response; stem cells; therapy outcome

Myeloproliferative neoplasms (MPNs) such as polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) often carry JAK2(V617F), MPL(W515L) and mutations in calreticulin (CALRmut). These aberrations may be accompanied by mutations in TET2, ASXL1, DNMT3A, EZH2, and other genes further complicating utilization of MPNs genetic/epigenetic signatures as potential biomarkers for therapeutic decisions. Current treatment options for MPNs include myelosuppressive therapy in the form of hydroxyurea and JAK1/2 inhibitor QAK1/2i) ruxolitinib. MPNs can have prolonged chronic phases, but may eventually accelerate and transform into a secondary acute myeloid leukemia that is ultimately fatal. Therefore, it is imperative to generate new therapies that alone or in combination with already approved drugs could potentially extend the complete remission time and/or be used in patients which progressed to the malignant stage. Since all 3 "main" mutations [JAK2 (V617F), CALR(del52), and MPL(W515L)] were found in MPN stem cells (MPNSCs) these cells must be eliminated in order to improve therapeutic outcome. Unfortunately, the potential therapeutic approaches against MPNSCs are limited. MPN cells, including MPNSCs accumulate potentially lethal DNA double-strand breaks (DSBs), which are repaired by two major mechanisms, BRCA-mediated homologous recombination (HR) and DNA-PK -mediated non-homologous end-joining (D-NHEJ). HR and D-NHEJ repair DSBs in proliferating cells and D-NHEJ plays a major role in quiescent cells. PARP1 -dependent back-up NHEJ (B-NHEJ) serves as back-up in both proliferating and quiescent cells. Cancer-specific defects in DSB repair create the opportunity to employ synthetic lethality, e.g. elimination of BRCA1/2-mutated cancer cells by PARP1 inhibitor (PARP1i). However, BRCA1/2 mutations are rare in MPNs. We hypothesize that MPN-inducing mutations are prognostic biomarkers of therapeutic synthetic lethality triggered by DNA repair inhibitors. We will determine if specific MPN-inducing mutation(s) (biomarkers) predispose quiescent and/or proliferating MPN stem and progenitor cells from individual patients to PARP1i-induced synthetic lethality combined with the inhibitors of HR and D-NHEJ (Aim #1) or with JAK2i (Aim #2). We will also employ murine knockin/knockout models of MPNs and primary MPN xenografts in humanized immunodeficient mice to determine if DNA repair inhibitors and/or JAK2i exert anti-MPN synthetic lethal effect in pre-clinical settings (Aim #3).

脊髓增生性肿瘤（MPN），例如多余的Vera（PV），必需血小板血症 （ET），一级骨髓纤维化（PMF）经常携带JAK2（V617F），MPL（W515L）和突变 钙网蛋白（calrmut）。这些畸变可能伴随着TET2，ASXL1中的突变 DNMT3A，EZH2和其他基因进一步使MPN遗传/表观遗传学的利用变得复杂 签名是治疗决定的潜在生物标志物。 MPN的当前治疗选择包括以骨髓性疗法的形式 羟基脲和JAK1/2抑制剂QAK1/2I）ruxolitinib。 MPN可以延长慢性相， 但最终可能会加速并转变为次要急性髓样白血病 最终致命。因此，必须单独或组合产生新的疗法 有了已经批准的药物，可能会延长完整的缓解时间和/或使用 在发展为恶性阶段的患者中。由于所有3个“主要”突变[JAK2（V617F），所以 在MPN干细胞（MPNSC）中发现CALR（DEL52）和MPL（W515L）]这些细胞必须为 消除以改善治疗结果。不幸的是，潜在的治疗 针对MPNSC的方法有限。 MPN细胞，包括MPNSC，积累了潜在的致命DNA双链断裂（DSB）， 通过两种主要机制修复，BRCA介导的同源重组（HR）和 DNA-PK介导的非同源最终连接（D-NHEJ）。 HR和D-NHEJ修复DSB 增殖细胞和D-NHEJ在静态细胞中起主要作用。 parp1依赖性备份NHEJ （B-NHEJ）在增殖和静态细胞中都可以作为备份。 DSB修复中的癌症特异性缺陷创造了使用合成致死性的机会，例如 通过PARP1抑制剂（PARP1I）消除BRCA1/2突变的癌细胞。但是，BRCA1/2 MPN中很少发生突变。我们假设诱导MPN的突变是预后的 DNA修复抑制剂触发的治疗合成致死性的生物标志物。 我们将确定特定的MPN诱导突变（S）（生物标志物）易感性静止 和/或从单个患者到PARP1I诱导的MPN茎和祖细胞增殖 合成致死性与HR和D-NHEJ（AIM＃1）或JAK2I（AIM＃2）的抑制剂结合在一起。 我们还将在MPN和主要MPN异种移植物中采用鼠敲击/敲除模型 人性化的免疫缺陷小鼠以确定DNA修复抑制剂和/或JAK2I是否发挥抗MPN 临床前环境中的合成致命作用（AIM＃3）。