Preclinical Development of First-in-Class GSTO1 Degraders for Colorectal Cancer

首创的结直肠癌 GSTO1 降解剂的临床前开发

• 批准号: 10675586
• 负责人: NOURI NEAMATI
• 金额: $ 53.14万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-02 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675586
• 关键词: 3-Dimensional; Bioinformatics; Bromouridine sequencing; CRISPR/Cas technology; Cell Adhesion Molecules; Cell Line; Cell Survival; Cells; Clinical; Clinical Research; Colorectal Cancer; Complex; Coupled; Critical Pathways; Data; Down-Regulation; Drug Combinations; Drug Targeting; Drug resistance; Engineering; Fluorouracil; Future; GSTO1 gene; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Genetic; Genetic Transcription; Immune checkpoint inhibitor; Immune response; Immunotherapy; Impairment; In Vitro; Innate Immune Response; Interferon Type I; Interferons; Interleukin-1 beta; Knock-out; Link; Malignant Neoplasms; Mediating; Microsomes; Mission; Modeling; Names; Natural Immunity; Neoplasm Metastasis; Oncology; Pathway interactions; Patients; Protein Isoforms; Proteomics; Public Health; Regulation; Risk Factors; Role; Series; Small Interfering RNA; Structure; Testing; Therapeutic; Thromboplastin; Time; Tumor Promotion; United States National Institutes of Health; Work; anti-PD1 antibodies; cancer cell; chemotherapy; clinical candidate; clinical development; colorectal cancer treatment; design; druggable target; efficacy study; immune activation; in vivo; in vivo Model; inhibitor; innate immune sensing; insight; irinotecan; lead optimization; monocyte; multiple omics; neoplastic cell; novel; novel anticancer drug; novel therapeutics; overexpression; oxaliplatin; patient derived xenograft model; pharmacodynamic biomarker; pharmacologic; preclinical development; protein expression; prototype; rational design; response; simulation; small hairpin RNA; small molecule; small molecule inhibitor; synergism; therapeutic target; tool; transcriptome sequencing; tumor; tumor growth; tumor progression; tumorigenesis

Glutathione S-transferase omega 1 (GSTO1) is an atypical GST isoform that is overexpressed in several cancers and has been implicated in drug resistance. Currently, no small-molecule drug targeting GSTO1 is under clinical development. Using genetic tools, including extensive bioinformatics analysis coupled with siRNA, shRNA, proteomics, and CRISPR/Cas9 technologies; and pharmacologic small-molecule inhibitors and degraders, we have validated GSTO1 as an impactful druggable target in oncology. Previously, we identified C1-27 as a potent GSTO1 inhibitor that shows efficacy against cancer cells in both in vitro and in vivo models. We also synthesized and tested the very first GSTO1 PROTAC confirming GSTO1 degradation in vitro. Through transcriptional profiling using Bru-seq and RNA-seq coupled with proteomics, we uncovered novel pharmacodynamic markers and cellular pathways critical for oncogenesis regulated by GSTO1. Taken together, our findings validate GSTO1 as an important drug target for cancer therapeutics and C1-27 as a potent and validated prototype inhibitor. Previously, we solved the crystal structure of C1-27 (IC50 = 31 nM) and other potent inhibitors in complex with GSTO1. Our most recent lead optimization campaign using 6 different co-crystal structures resulted in the most potent GSTO1 inhibitor (IC50 = 0.22 ± 0.02 nM) known to date. Our CRISPR/Cas9 GSTO1 knockout (KO) cell lines do not form tumors or display tumor growth delay in vivo and form smaller 3D spheroids in vitro. Through multi-omics studies in GSTO1 KO cells, we found a strong positive correlation with cell adhesion molecules and interferon response pathways, and a strong negative correlation with Myc transcriptional signature. Importantly, we also identified several clinically used chemotherapies showing significant synthetic lethality with loss or inhibition of GSTO1. We discovered that tissue factor (gene name, F3) transcription and protein expression are downregulated in response to GSTO1 KO and C1-27 treatment, further implicating a role for GSTO1 in the innate immune response. In summary, our results implicate GSTO1 as a therapeutic target in cancer and offer new mechanistic insights into its significant role in cancer progression. Importantly, our results show for the first time that inhibition of GSTO1 can activate immune responses and downregulate F3. We hypothesize that inhibiting GSTO1 will have a two-pronged effect on tumor cells: (A) impair cancer cell survival by reducing Myc transcriptional signature and F3 downregulation, and (B) enhance immune responses through interferon- mediated innate immune sensing of cancers. We further hypothesize that GSTO1 inhibitors will sensitize cancer cells to select chemotherapy and immunotherapy. We will test these hypotheses through the following three specific aims. Aim 1: Elucidate the functions of GSTO1 in enhancing immune response through activation of type-I interferon and reducing tumor cell viability through F3 and Myc downregulation. Aim 2: Optimize GSTO1 degraders based on co-crystal structures to select a clinical candidate. Aim 3: Assess in vivo efficacy of promising novel GSTO1 degraders in PDX and syngeneic models of CRC as single agents and in combination.

谷胱甘肽S-转移酶ω 1（GSTO 1）是一种非典型的GST亚型，在几种癌症中过表达 并且与抗药性有关目前，尚无靶向GSTO 1的小分子药物进入临床 发展使用遗传工具，包括广泛的生物信息学分析加上siRNA，shRNA， 蛋白质组学和CRISPR/Cas9技术;以及药理学小分子抑制剂和降解剂，我们 已经验证了GSTO 1作为肿瘤学中有影响力的药物靶点。之前，我们发现C1-27是一种有效的 GSTO 1抑制剂，在体外和体内模型中都显示出对癌细胞的功效。我们还合成 并测试了第一个GSTO 1 PROTAC，证实了GSTO 1的体外降解。通过转录 使用Bru-seq和RNA-seq结合蛋白质组学进行分析，我们发现了新的药效学标志物 和细胞通路的关键肿瘤发生的GSTO 1调节。综上所述，我们的研究结果验证了GSTO 1 作为癌症治疗的重要药物靶标，C1-27作为有效且经验证的原型抑制剂。 以前，我们解决了C1-27（IC 50 = 31 nM）和其他有效抑制剂的晶体结构， GSTO 1.我们最近的铅优化活动使用6种不同的共晶结构， 迄今已知的有效GSTO 1抑制剂（IC 50 = 0.22 ± 0.02 nM）。我们的CRISPR/Cas9 GSTO 1敲除（KO）细胞 线在体内不形成肿瘤或显示肿瘤生长延迟，并且在体外形成较小的3D球状体。通过 在GSTO 1 KO细胞的多组学研究中，我们发现与细胞粘附分子和 干扰素反应途径，并与Myc转录签名强负相关。重要的是， 我们还确定了几种临床使用的化疗药物，显示出显著的合成致死性， 抑制GST 0 1。我们发现，组织因子（基因名称，F3）转录和蛋白质表达是 GSTO 1 KO和C1-27处理后表达下调，进一步表明GSTO 1在先天性免疫缺陷中的作用。 免疫反应总之，我们的研究结果暗示GSTO 1作为癌症的治疗靶点，并提供了新的 其在癌症进展中的重要作用的机制见解。重要的是，我们的研究结果首次表明， GSTO 1的抑制可以激活免疫反应并下调F3。我们假设抑制 GSTO 1对肿瘤细胞有两方面的影响：（A）通过减少Myc， 转录标签和F3下调，和（B）通过干扰素- 介导的癌症先天免疫感应。我们进一步假设GSTO 1抑制剂会使癌症敏感， 选择化疗和免疫疗法。我们将通过以下三个方面来检验这些假设 明确的目标。目的1：阐明GSTO 1通过激活细胞因子，增强免疫应答的功能。 I型干扰素和通过F3和Myc下调降低肿瘤细胞活力。目标2：优化GSTO 1 基于共晶结构的降解剂来选择临床候选物。目的3：评估有希望的 在PDX和CRC的同基因模型中作为单一药剂和组合的新型GST 0 1降解剂。