Targeting the Sam68-stimulated PARP1 activation for cancer treatment

针对 Sam68 刺激的 PARP1 激活进行癌症治疗

• 批准号: 10163145
• 负责人: Fengyi Wan
• 金额: $ 13.54万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 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 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10163145
• 关键词: Affect; Anemia; Animals; Attenuated; Basic Science; Biochemical; Biological Assay; Biophysics; Bos taurus PARP protein; Cancerous; Categories; Cell Death; Cell physiology; Cells; Cellular Assay; Chemicals; Clinical; Coenzymes; Colon; Colon Carcinoma; Colonic Neoplasms; Competitive Binding; DNA Damage; DNA Repair; DNA lesion; Development; Disease; Enzyme-Linked Immunosorbent Assay; Fatigue; Health; Human; In Vitro; Interruption; Intervention; Knock-out; Laboratories; Lead; Libraries; Malignant Neoplasms; Mitosis; Modality; Modeling; Molecular; Mus; Myelosuppression; Nausea; Neutropenia; Niacinamide; Nicotinamide adenine dinucleotide; Normal Cell; Observational Study; Pharmacology; Play; Poly(ADP-ribose) Polymerases; Production; Proteins; Radiation therapy; Regulation; Research; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Specificity; Therapeutic; Tissues; Toxic effect; Tumor Burden; Tumor Tissue; Validation; base; cancer cell; cancer survival; cancer therapy; cancer type; colon cancer cell line; experimental study; follow-up; gamma irradiation; gastrointestinal; genotoxicity; high throughput screening; imaging modality; improved; inhibitor/antagonist; innovation; interdisciplinary approach; knock-down; millisecond; mouse model; negative affect; novel; novel strategies; public health relevance; recruit; repaired; response; screening; small molecule; small molecule inhibitor; small molecule libraries; therapeutic target

Project Summary The inhibition of poly(ADP-ribose) polymerase 1 (PARP1) protein has emerged as a promising therapeutic treatment for several types of cancers. However, the current classes of PARP1 inhibitors are all derivatives of nicotinamide, based on the strategy of competitive binding against nicotinamide. Because the PARP1-catalyzed PARylation is critical for many cellular processes, therapeutic targeting of PARP1 in cancerous cells with current classes of PARP1 inhibitors could negatively affect many cell functions in normal cells. There is an urgent clinical need for improving the inhibitor specificity for PARP1 and lowering the off- target effects and toxicity for better therapeutic value and modalities. However, the rapid recruitment of PARP1 to sites of DNA lesions (within milliseconds) and the vigorously synthesized PAR chains make it difficult to elucidate the precise stimulatory and regulatory mechanisms on PARP1 activation. We recently discovered that Src-associated-substrate-during-mitosis-of-68kDa (Sam68) plays an important role in stimulating the DNA damage-specific PARP1 activation, which suggests that targeting the Sam68-stimulated PARP1 activation could be a novel strategy to develop a new category of PARP1 inhibitors. The objective of our research is to identify specific molecule inhibitors of the Sam68-stimulated PARP1 activation via a high throughput screening, and our hypothesis is that these inhibitors would alleviate DNA damage-triggered PARP1 activation thus being potential pharmacological agents for treating cancers. We have developed a novel ELISA-based assay suitable for screening of small molecule libraries. In this proposed research, we will conduct high throughput screening to identify small molecule compounds inhibiting the Sam68-stimulated PARP1 activation. We will employ interdisciplinary approach combining biochemical, biophysical, and molecular experiments, cellular assays, and imaging methods to validate the screening hits and to identify high quality small molecule inhibitors of Sam68-PARP1 interaction. Activity of the most potent compounds will be evaluated in a panel of cell-based and mouse model experiments to assess their capability to inhibit the survival and development of colon cancer cells. Our project represents a very innovative strategy to inhibit PARP1 activity through blocking the Sam68-conferred stimulatory mechanism. At the conclusion of these studies, we expect to identify highly valuable compounds that can serve as chemical probes suitable for further development into potent PARP1 inhibitors for cancer therapeutics. Additionally, discovery of these inhibitors will greatly facilitate our research on the regulation and function of the Sam68-stimulated PARP1 activation under normal and disease conditions.

项目摘要 聚（ADP-核糖）聚合酶1（PARP1）蛋白的抑制作用已成为有前途的 几种类型的癌症的治疗治疗。但是，当前的PARP1抑制剂都是 烟酰胺的衍生物基于与烟酰胺的竞争性结合策略。因为 PARP1催化的核化对许多细胞过程至关重要，PARP1的治疗靶向 具有当前类PARP1抑制剂类别的癌细胞可能会对正常的许多细胞功能产生负面影响 细胞。紧急临床需要提高抑制剂特异性的PARP1并降低非抑制剂特异性 目标效应和毒性，以提高治疗价值和方式。但是，快速招募 PARP1到DNA病变（毫秒内）和剧烈合成的PAR链的位置 难以阐明PARP1激活的精确刺激和调节机制。我们最近 发现68kDa（SAM68）在 刺激DNA损伤特异性PARP1激活，这表明针对SAM68刺激的 PARP1激活可能是开发新类别PARP1抑制剂的新型策略。目的 我们的研究是通过高 吞吐量筛选，我们的假设是这些抑制剂会减轻受DNA损伤的触发 PARP1激活是治疗癌症的潜在药理剂。我们已经开发了 基于ELISA的新型测定法，适合筛选小分子文库。在这项拟议的研究中，我们将 进行高通量筛选以鉴定抑制SAM68刺激的小分子化合物 PARP1激活。我们将采用跨学科方法，结合生化，生物物理和 分子实验，细胞测定和成像方法，以验证筛选并鉴定 SAM68-PARP1相互作用的高质量小分子抑制剂。最有效化合物的活动将 在基于细胞和小鼠模型实验的小组中进行评估，以评估其抑制的能力 结肠癌细胞的生存和发育。我们的项目代表了一种抑制的非常创新的策略 PARP1通过阻断SAM68授予的刺激机制的活性。在这些结束时 研究，我们期望确定高价的化合物，可以用作适合的化学探针 进一步发展为癌症治疗剂有效的PARP1抑制剂。另外，发现这些 抑制剂将极大地促进我们对SAM68刺激的PARP1的调节和功能的研究 在正常和疾病条件下的激活。