Cause and therapeutic impact of DNA-protein crosslink repair defect in myeloid leukemias

髓系白血病 DNA-蛋白质交联修复缺陷的原因和治疗影响

基本信息

批准号：
10296087
负责人：
SCOTT H KAUFMANN
金额：
$ 36.37万
依托单位：
MAYO CLINIC ROCHESTER
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：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10296087
关键词：
AML/MDS Acute Lymphocytic Leukemia Acute Myelocytic Leukemia Affect Age Anthracycline Antibodies Azacitidine Biochemical Camptothecin Cancer Therapy Evaluation Program Case Fatality Rates Cell Line Cells Chronic Myelomonocytic Leukemia Clinic Clinical Complex Cytotoxic agent DNA DNA Damage DNA Methyltransferase Inhibitor DNA Modification Methylases DNA Topoisomerases DNA lesion DNA-protein crosslink Daunorubicin Decitabine Defect Development Disease Drug usage Enzymes Etoposide Excision Exhibits Exposure to Formaldehyde Generations Hematopoietic Hematopoietic Stem Cell Transplantation Hemorrhagic Thrombocythemia Human Immunologics In Vitro Knock-out Lead Lesion Leukemic Cell Lymphoid Cell Malignant - descriptor Marrow Measures Metabolism Metalloproteases Mitoxantrone Myeloid Cells Myeloid Leukemia Myeloproliferative disease Neoplasms Nuclear Participant Pathway interactions Patients Peptide Hydrolases Pharmaceutical Preparations Pharmacotherapy Phase Play Poison Polycythemia Vera Process Reagent Regimen Role Sampling Serine Protease Specimen Stabilizing Agents Syndrome TOP1 gene TOP2A gene Testing Therapeutic Tissues Topoisomerase Topotecan acute myeloid leukemia cell antileukemic agent base cell type chromatin protein comorbidity covalent bond crosslink design drug sensitivity high reward high risk histone demethylase improved improved outcome in vivo in vivo Model insight leukemia nuclease phase II trial phosphoric diester hydrolase progenitor repaired research clinical testing response targeted agent targeted treatment

项目摘要

ABSTRACT Acute myeloid leukemias (AMLs) are a genetically heterogeneous group of clonal hematopoietic disorders characterized by accumulation of immature non-lymphoid marrow progenitors. While there have been notable therapeutic advances over the past 5 years, many AML subtypes continue to have case fatality rates of >50%. Despite the introduction of a number of targeted therapies, conventional cytotoxic drugs – alone or in combination with the targeted agents – remain the mainstay of AML therapy. Among the conventional cytotoxic drugs used to treat AML, several act by increasing unique types of DNA lesions known as DNA-protein crosslinks (DPCs). In particular, topoisomerase poisons increase the number of DPCs containing TOP2 (daunorubicin, mitoxantrone, etoposide) or TOP1 (topotecan) covalently bound to DNA. In addition, the hypomethylating agents decitabine and azacitidine increase the number of DPCs containing DNA methyltransferases covalently bound to DNA. The mechanisms involved in DPC repair are at present incompletely understood. To facilitate the further development of topotecan and other TOP1 poisons, as well contribute to the study of TOP1-containing DPCs, we have generated an antibody that specifically recognizes TOP1-DNA covalent complexes (TOP1ccs). Using this antibody, we have shown that the nuclear metalloproteinase SPARTAN and the serine protease FAM111A, acting upstream of the phosphodiesterase TDP1, play important roles in the repair of TOP1ccs in some tissues. Importantly, loss of SPARTAN, FAM111A or TDP1 leads to accumulation of TOP1ccs in the absence of drug treatment as well as enhanced sensitivity to the prototypic TOP1 poison camptothecin. More recently, we have also observed that a variety of malignant myeloid cells, including AML cell lines and primary AML specimens, contain readily detectable TOP1ccs in the absence of drug treatment and are slow to repair TOP1ccs upon exposure to the TOP1 poison topotecan. In contrast, the vast majority of tissues, including normal and malignant lymphoid cells as well as normal marrow, contain very few TOP1ccs in the absence of drug treatment. These results lead to the hypothesis that many myeloid neoplasms have previously unsuspected defects in TOP1cc repair that might affect their therapeutic sensitivity. To test this hypothesis, we now propose to define the biochemical basis for the constitutive increase in TOP1ccs in myeloid neoplasms (Aim 1), examine the impact of low TOP1cc repair on leukemia cell sensitivity to agents that stabilize DPCs (Aim 2), and assess the relationship between TOP1cc levels (constitutive and drug-induced) and clinical response of myeloid neoplasms to a topotecan-containing regimen currently undergoing NCI-sponsored phase II clinical testing in high risk AML (Aim 3). Collectively, these studies will provide important new insight into a previously unsuspected DPC repair defect in myeloid malignancies and begin to determine whether this repair defect has therapeutic implications that can be used to guide improvements in AML therapy.

抽象的急性髓系白血病 (AML) 是一组具有遗传异质性的克隆性造血系统疾病其特征是未成熟的非淋巴骨髓祖细胞的积累，同时也存在值得注意的情况。过去 5 年治疗取得了进展，许多 AML 亚型的病死率仍然超过 50%。尽管引入了许多靶向疗法，但传统的细胞毒性药物——单独使用或联合使用与靶向药物的联合治疗仍然是 AML 传统细胞毒性治疗的支柱。用于治疗 AML 的几种药物通过增加称为 DNA 蛋白质的独特类型 DNA 损伤来发挥作用特别是，拓扑异构酶毒物会增加含有 TOP2 的 DPC 的数量。（柔红霉素、米托蒽醌、依托泊苷）或 TOP1（拓扑替康）与 DNA 共价结合。去甲基化药物地西他滨和阿扎胞苷增加含有 DNA 的 DPC 数量甲基转移酶与 DNA 共价结合，目前涉及 DPC 修复的机制。尚不完全了解，以促进拓扑替康和其他 TOP1 毒物的进一步开发。为了对含有 TOP1 的 DPC 的研究做出贡献，我们已经生成了一种特异性识别使用该抗体，我们显示了 TOP1-DNA 共价复合物 (TOP1ccs)。金属蛋白酶 SPARTAN 和丝氨酸蛋白酶 FAM111A，作用于磷酸二酯酶的上游 TDP1 在某些组织中 TOP1cc 的修复中发挥重要作用，重要的是 SPARTAN、FAM111A 的丢失。或 TDP1 在没有药物治疗的情况下导致 TOP1cc 的积累以及对药物敏感性的增强原型TOP1毒药喜树碱最近，我们还观察到多种恶性。骨髓细胞，包括 AML 细胞系和原代 AML 样本，在细胞中含有易于检测的 TOP1cc 缺乏药物治疗，并且在暴露于 TOP1 毒药拓扑替康后修复 TOP1cc 缓慢。相比之下，绝大多数组织，包括正常和恶性淋巴细胞以及正常骨髓，在没有药物治疗的情况下，TOP1ccs 含量非常少。这些结果导致了这样的假设：许多。骨髓肿瘤在 TOP1cc 修复中存在先前未被怀疑的缺陷，这可能会影响它们的为了检验这一假设，我们现在建议定义其生化基础。骨髓肿瘤中 TOP1cc 的组成性增加（目标 1），检查低 TOP1cc 修复对白血病细胞对稳定 DPC 的药物的敏感性（目标 2），并评估 TOP1cc 之间的关系骨髓肿瘤对含拓扑替康的水平（组成型和药物诱导型）和临床反应目前正在接受 NCI 赞助的高风险 AML 的 II 期临床测试（目标 3）。这些研究将为先前未曾怀疑的骨髓中 DPC 修复缺陷提供重要的新见解恶性肿瘤并开始确定这种修复缺陷是否具有可以使用的治疗意义指导 AML 治疗的改进。