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

基本信息

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
项目类别：
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

项目摘要

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 omega 1 (GSTO1) 是一种非典型 GST 亚型，在多种癌症中过度表达目前还没有针对GSTO1的小分子药物进入临床。使用遗传工具，包括结合 siRNA、shRNA 的广泛生物信息学分析，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，我们已验证 GSTO1 是肿瘤学中有效的药物靶点此前，我们将 C1-27 确定为有效的药物靶点。我们还合成了在体外和体内模型中对癌细胞有效的 GSTO1 抑制剂。并测试了第一个 GSTO1 PROTAC，确认 GSTO1 通过转录降解。使用 Bru-seq 和 RNA-seq 与蛋白质组学相结合进行分析，我们发现了新的药效标记物以及对 GSTO1 调节的肿瘤发生至关重要的细胞通路综上所述，我们的研究结果验证了 GSTO1。作为癌症治疗的重要药物靶点，C1-27 作为有效且经过验证的原型抑制剂。之前，我们解析了 C1-27 (IC50 = 31 nM) 和其他有效抑制剂与我们最近使用 6 种不同的共晶结构进行的先导化合物优化活动产生了最多的结果。迄今为止已知的有效 GSTO1 抑制剂 (IC50 = 0.22 ± 0.02 nM) 我们的 CRISPR/Cas9 GSTO1 敲除 (KO) 细胞。细胞系在体内不会形成肿瘤或表现出肿瘤生长延迟，并且在体外会形成较小的 3D 球体。在 GSTO1 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，并且与 Myc 转录特征呈强负相关。我们还确定了几种临床使用的化疗药物，它们显示出显着的合成致死率，并伴有损失或我们发现组织因子（基因名称，F3）转录和蛋白质表达受到抑制。 GSTO1 KO 和 C1-27 治疗后下调，进一步暗示 GSTO1 在先天性总之，我们的结果表明 GSTO1 作为癌症的治疗靶点并提供了新的思路。重要的是，我们的结果首次表明其在癌症进展中的重要作用。抑制 GSTO1 可以激活免疫反应并下调 F3。 GSTO1将对肿瘤细胞产生双管齐下的影响：(A)通过减少Myc来损害癌细胞的存活转录特征和 F3 下调，以及 (B) 通过干扰素增强免疫反应我们进一步发现 GSTO1 抑制剂会使癌症变得敏感。我们将通过以下三个方面来检验这些假设。具体目标 1：阐明 GSTO1 通过激活增强免疫反应的功能。 I 型干扰素并通过下调 F3 和 Myc 降低肿瘤细胞活力目标 2：优化 GSTO1。目的 3：评估有前景的体内功效。新型 GSTO1 降解剂在 PDX 和 CRC 同基因模型中作为单一药物或组合使用。