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: 10438886
• 负责人: SCOTT H KAUFMANN
• 金额: $ 35.64万
• 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/10438886
• 关键词: 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; Dysmyelopoietic Syndromes; 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)是一组遗传异质性的克隆性造血疾病 以聚集未成熟的非淋巴系骨髓祖细胞为特征。虽然有值得注意的是 治疗进展在过去5年中，许多AML亚型的病死率仍高达50%。 尽管引入了一些靶向治疗，但传统的细胞毒性药物--单独或 联合靶向药物--仍然是急性髓系白血病治疗的主要手段。在传统的细胞毒中 用于治疗AML的药物中，有几种是通过增加被称为DNA-蛋白质的独特类型的DNA损伤来发挥作用的 交叉链接(DPC)。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。此外， 去甲基化药物地西他滨和阿扎替丁增加含有DNA的DPC的数量 甲基转移酶与DNA共价结合。目前，参与DPC修复的机制有 不完全理解。促进拓扑替康和其他TOP1毒素的进一步发展 对含有TOP1的DPC的研究做出了贡献，我们已经产生了一种抗体，可以特异性地识别 Top1-DNA共价复合体(Top1ccs)。使用这种抗体，我们已经证明了核 作用于磷酸二酯酶上游的金属蛋白酶Spartan和丝氨酸蛋白酶FAM111a Tdp1在一些组织中Top1ccs的修复中起重要作用。重要的是，失去斯巴达人，FAM111A 或Tdp1导致Top1ccs在没有药物治疗的情况下积聚，以及对 原型TOP1毒药喜树碱。最近，我们还观察到各种恶性 髓系细胞，包括AML细胞系和原代AML标本，在 缺乏药物治疗，在接触TOP1毒物拓扑替康后修复Top1ccs速度较慢。在……里面 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，含有极少的Top1ccs。这些结果导致了这样的假设：许多 髓系肿瘤在Top1cc修复中有先前未被怀疑的缺陷，这可能会影响他们的 治疗敏感度。为了检验这一假设，我们现在建议定义 髓系肿瘤中Top1ccs的结构性增加(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与 髓系肿瘤对含拓扑替康的药物浓度(结构性和药物诱导)和临床反应 目前正在接受NCI赞助的高危AML第二阶段临床测试的方案(AIM 3)。总而言之， 这些研究将为一种以前未被怀疑的髓系DPC修复缺陷提供重要的新见解 并开始确定这种修复缺陷是否具有可以使用的治疗意义 以指导急性髓系白血病治疗的改进。