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

• 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
• 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

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.

摘要 急性髓系白血病（AML）是一组遗传异质性克隆性造血系统疾病 其特征在于未成熟的非淋巴样骨髓祖细胞的积累。虽然有一些值得注意的 尽管在过去5年的治疗进展中，许多AML亚型的病死率仍> 50%。 尽管引入了许多靶向疗法，但常规细胞毒性药物-单独或联合 与靶向药物联合治疗仍然是AML治疗的主要手段。在传统的细胞毒性药物中， 用于治疗AML的药物中，有几种通过增加称为DNA蛋白的独特类型的DNA损伤来起作用。 交联（DPC）。特别地，拓扑异构酶毒物增加含有TOP2的DPC的数量， （柔红霉素、米托蒽醌、依托泊苷）或TOP 1（拓扑替康）共价结合到DNA。此外该 低甲基化剂地西他滨和阿扎胞苷增加含有DNA的DPC的数量， 与DNA共价结合的甲基转移酶。目前DPC修复所涉及的机制是 不完全理解。为促进拓扑替康及其他TOP1类药物的进一步开发， 为了有助于研究含有TOP1的DPC，我们已经产生了一种特异性识别 TOP1-DNA共价复合物（TOP1 ccs）。使用这种抗体，我们已经证明， 金属蛋白酶SPARTAN和丝氨酸蛋白酶FAM 111 A，作用于磷酸二酯酶的上游 在某些组织中，TDP 1在TOP1 ccs的修复中起重要作用。重要的是，失去了斯巴达，FAM 111 A 或TDP 1导致在没有药物治疗的情况下TOP1 ccs的积累以及对 原型TOP 1有毒喜树碱。最近，我们还观察到， 骨髓细胞，包括AML细胞系和原发性AML标本，在骨髓细胞中含有易于检测的TOP1 ccs。 缺乏药物治疗，并且在暴露于TOP1毒物拓扑替康后修复TOP1 ccs缓慢。在 相反，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，TOP1 ccs很少。这些结果导致了一种假设， 髓系肿瘤在TOP1 cc修复中存在先前未被怀疑的缺陷，这可能影响其 治疗敏感性为了验证这一假设，我们现在提出定义的生化基础， 骨髓肿瘤中TOP1 ccs的组成性增加（目的1），检查低TOP1 cc修复对 白血病细胞对稳定DPC的药物的敏感性（目的2），并评估TOP1 cc 骨髓肿瘤对含托泊替康的药物的水平（组成性和药物诱导性）和临床反应 目前正在接受NCI申办的高风险AML II期临床试验的方案（Aim 3）。总的来说， 这些研究将为以前未被怀疑的骨髓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治疗意义， 以指导AML治疗的改进。