PROJECT 2

项目2

• 批准号: 9804092
• 负责人: Daniel A Harki
• 金额: $ 36.09万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-09 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9804092
• 关键词: Achievement; Address; Advanced Development; Animal Model; Animals; Antibody Formation; Antiviral Agents; Automobile Driving; BRCA1 gene; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Biophysics; Breast; Breast Cancer Cell; Breast Cancer Model; Cells; Cellular Assay; Chemicals; Clinical; Collaborations; Complex; Crystallization; Cytosine; DNA; DNA Binding; DNA Sequence; Deaminase; Deamination; Defect; Deoxycytidine; Development; Drug resistance; Enzyme Inhibition; Enzymes; Estrogen receptor positive; Evolution; Foundations; Future; Goals; Graph; Inherited; Lead; Ligand Binding; Ligands; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metastatic breast cancer; Methods; Modeling; Modification; Molecular; Mutagenesis; Mutate; Mutation; Neoplasm Metastasis; Nucleic Acid Probes; Nucleic Acids; Nucleotides; Oligonucleotides; Outcome; PIK3CA gene; Physiological; Positioning Attribute; Primary Neoplasm; Process; Proteins; RNA; Reagent; Reporting; Research; Resistance; Roentgen Rays; Role; Services; Single-Stranded DNA; Structure; Technology; Testing; The Cancer Genome Atlas; Therapeutic; Treatment Failure; Uracil; Variant; Virus Diseases; Work; base; clinical translation; computational chemistry; design; drug development; drug discovery; experimental study; feature detection; forging; homologous recombination; improved; in vivo; inhibitor/antagonist; innovation; malignant breast neoplasm; molecular recognition; mouse model; multidisciplinary; novel; novel strategies; overexpression; preference; prevent; programs; small molecule; small molecule inhibitor; structural biology; therapeutic development; therapy outcome; tumor

PROJECT 2 – CHEMICAL BIOLOGY OF DNA DEAMINASES IN BREAST CANCER ABSTRACT APOBEC enzymes are single-stranded DNA cytosine-to-uracil deaminases that normally protect cells from viral infections. However, APOBEC3B (A3B) has been implicated in mutations in breast cancer that drive tumor evolution and contribute to the development of drug resistance and, ultimately, therapy failure. A3B is overexpressed in over half of all estrogen receptor (ER)-positive breast tumors, the most common type, and is associated with poor overall survival. Our Program has shown that inhibition of A3B-mediated tumor evolution improves therapy outcomes in a mouse model of ER-positive breast cancer. Our Program’s unifying hypothesis is that A3B inhibition will prevent a large proportion of new mutations in ER-positive breast cancer, thereby improving the durability of current treatments and resulting in better overall outcomes. To address this hypothesis our Program is focused on understanding the biology of A3B in breast cancer cells (Project 1); developing innovative nucleic acid probes to molecularly characterize how A3B engages DNA substrates and small molecules to inhibit A3B-catalyzed breast cancer mutations (Project 2); and generating A3B x-ray structures with nucleic acids, small molecules, and protein ligands to understand the structural basis of A3B- mediated DNA mutagenesis and its inhibition to enable development of therapeutic compounds (Project 3). These activities will be supported by Service Cores for administration (Core A), animal models of A3B-driven breast cancer (Core B), computational chemistry and biophysics (Core C), and protein and antibody production (Core D). Project 2 – Chemical Biology of DNA Deaminases in Breast Cancer will lead the chemical probe discovery efforts by 1) synthesizing complex nucleic acid ligands for A3B to characterize how A3B discriminates 2¢-deoxycytidine from other nucleotides and to understand which nucleic acid features enable A3B to deaminate discrete DNA sequences, including its overall preference for binding DNA versus RNA; and 2) using complimentary technologies and approaches to develop first-in-class small molecule inhibitors of A3B that will be used in mechanistic cellular assays of A3B-driven breast cancer mutation (Project 1), structural biology studies to annotate A3B-ligand binding (Project 3), and therapeutic utility experiments in animal models of breast cancer (Core B). Our studies will be enabled by critical collaborations involving computational ligand design (Core C) and access to high-quality biological reagents for assays (Core D), and our advances will position our novel compounds for future therapeutic development. Potent, selective chemical probes of A3B with in vivo activity, as well as novel assays, are the major anticipated deliverables of Project 2. As such, Project 2 will be the center of chemical innovation for the Program, accelerating all Projects and contributing to the achievement of our overall research objectives.

项目2-乳腺癌中DNA脱氨酶的化学生物学 摘要 APOBEC酶是单链DNA胞嘧啶-尿嘧啶脱氨酶，其通常保护细胞免于 病毒感染然而，APOBEC3B（A3B）与乳腺癌中驱动肿瘤的突变有关。 进化，并有助于耐药性的发展，最终导致治疗失败。A3B是 在超过一半的雌激素受体（ER）阳性乳腺肿瘤中过表达，这是最常见的类型， 与总体生存率低有关。我们的项目表明，抑制A3B介导的肿瘤演变 改善ER阳性乳腺癌小鼠模型的治疗结果。我们的计划是统一的 假设A3B抑制将防止ER阳性乳腺癌中大部分新突变， 从而提高当前治疗的持久性并导致更好的总体结果。为了解决这个 假设我们的计划专注于了解乳腺癌细胞中A3B的生物学（项目1）; 开发创新的核酸探针，从分子上表征A3B如何与DNA底物结合， 抑制A3B催化的乳腺癌突变的小分子（项目2）; 与核酸，小分子和蛋白质配体的结构，以了解A3B的结构基础- 介导的DNA诱变及其抑制，从而能够开发治疗化合物（项目3）。 这些活动将得到管理服务核心（核心A）、A3B驱动的动物模型 乳腺癌（核心B）、计算化学和生物物理学（核心C）以及蛋白质和抗体生产 (Core D）。项目2-乳腺癌中DNA脱氨酶的化学生物学将领导化学探针 通过1）合成A3B的复杂核酸配体以表征A3B如何 将2 ′-脱氧胞苷与其他核苷酸区分开来，并了解哪些核酸特征能够 A3B使离散的DNA序列脱氨基，包括其结合DNA相对于RNA的总体偏好;和 2)利用互补技术和方法开发一流的A3B小分子抑制剂 将用于A3B驱动的乳腺癌突变的机械细胞测定（项目1），结构 注释A3B-配体结合的生物学研究（项目3），以及动物模型中的治疗效用实验 乳腺癌（核心B）。我们的研究将通过涉及计算配体的关键合作来实现 设计（核心C）和获得高质量的生物试剂进行测定（核心D），我们的进步将 为未来的治疗开发定位我们的新化合物。A3B的有效、选择性化学探针 具有体内活性的药物以及新的检测方法是项目2的主要预期交付成果。因此，在本发明中， 项目2将成为该计划的化学创新中心，加速所有项目，并为 实现我们的整体研究目标。