Preventing invasive prostate cancer

翻译后摘要：原位癌形成浸润性原发性前列腺癌（PCa）所需的细胞运动。抑制 运动性将阻止侵袭性癌症的形成。到目前为止，还不可能选择性地抑制 能动性我们发现了一个一流的特工KBU 2046它选择性地抑制运动， 一个广泛的治疗窗口，使我们发现迄今未知的调控途径，目标是那些 途径，在GLP级IND使能研究中表现良好，并开辟了新的高价值创新概念 用于癌症预防拦截。研究支持KBU 2046是有效癌症的假设 拦截剂，通过一种新的机制运作，它和它调节的途径将有 具有抑制侵袭性癌症发展的高潜力，而侵袭性癌症又具有致命潜力。 目标1.鉴定并验证KBU 2046的药理学结合位点。计算机模拟，生物物理和生物化学 研究表明，KBU 2046结合在HSP 90 β和CDC 37之间的蛋白质-蛋白质界面， 选择性地减少与Raf 1的结合。使用基于KBU 2046的光交联探针和蛋白水解 靶向嵌合体（PROTAC），结合化学蛋白质组学方法和生物信息学分析，研究 将鉴定和验证细胞环境中的KBU 2046结合位点。 目标二。表征Raf 1调节人前列腺细胞运动的信号通路。我们 显示KBU 2046降低Raf 1激活基序上ser 338的磷酸化， 运动是通过一种新的途径介导的，其中Raf 1与肌动蛋白结合蛋白L-形成复合物， 而质谱仪（MS）蛋白质组学/生物信息学分析证明了主要作用 对细胞运动蛋白质的研究研究将在有风险的人类前列腺细胞中检查这一途径，将使用Raf 1 活性/非活性构建体，类似地工程化LCP 1构建体，通过正交分析探测信号传导， 使用MS对蛋白质特征的影响进行分析，并将研究单个途径成员对细胞增殖的作用。 运动性和KBU 2046功效。 目标3.在临床相关的人类疾病模型中评估靶向运动。在RapidCaP转基因 Pten和p53共缺失可诱导原位→侵袭→转移的PCa。它支持治疗之前， 与原位病变发展后相比，使我们能够检查预防和拦截效果， 分别用于抑制侵袭性癌症的形成。利用我们前列腺快速尸检的原代细胞 程序（RAP），我们表现出机械相关的响应KBU 2046。原代细胞和产物 类器官提供了在临床相关模型中检查功效的独特机会。RapidCaP和RAP 将在分子和功能水平上研究组织以检查KBU 2046的作用。 冲击研究人员寻求推进细胞运动是一种重要癌症的创新概念 预防拦截目标。它适用于许多癌症类型。

Abstract: Cell motility is required for in situ cancer to form invasive primary prostate cancer (PCa). Inhibiting motility would intercept formation of invasive cancer. Until now it has not been possible to selectively inhibit motility. We have discovered a precision acting first-in-class agent, KBU2046. It selectively inhibits motility, has a wide therapeutic window, has led us to uncover heretofore unknown regulatory pathways, targets those pathways, performs well in GLP grade IND enabling studies, and opens up new high value innovative concepts for cancer prevention-interception. Studies support the hypothesis that KBU2046 is an effective cancer interception agent that operates through a novel mechanism, and that it and the pathways it modulates will have high potential for inhibiting the development of invasive cancer, which in turn has lethal potential. Aim 1. Identify and validate KBU2046’s pharmacologic binding site. In silico, biophysical and biochemical studies suggest that KBU2046 binds at the protein-protein interface between HSP90β and CDC37 and selectively decreases binding to Raf1. Using KBU2046-based photo-crosslinkable probes and proteolysis targeting chimeras (PROTACs), coupled to chemo-proteomic approaches and bioinformatic analysis, studies will identify and validate KBU2046 binding sites in the cellular milieu. Aim 2. Characterize the signaling pathway by which Raf1 regulates the motility of human prostate cells. We show KBU2046 decreases phosphorylation of ser338 on Raf1’s activation motif, that downstream regulation of motility is mediated through a novel pathway wherein Raf1 forms a complex with the actin binding protein, L- plastin (LCP1), while mass spectrometer (MS) proteomic / bioinformatic analysis demonstrates primary effects on cell motility proteins. Studies will examine this pathway in at risk human prostate cells, will use Raf1 active/inactive constructs, similarly engineer LCP1 constructs, probe signaling through orthogonal analytics, profile effects on protein signature using MS, and will examine the role of individual pathway members on cell motility and KBU2046 efficacy. Aim 3. Evaluate targeting motiliy in clinically relevant models of human disease. In the RapidCaP transgenic model, codeletion of Pten and p53 induces in situ→invasive→metastatic PCa. It supports treatment before versus after development of in situ lesions, allowing us to examine prevention and interception efficacy, respectively, for inhibiting formation of invasive cancer. Using primary cells from our prostate rapid autopsy program (RAP), we show mechanistically relevant responsiveness to KBU2046. Primary cells and resultant organoids provide a unique opportunity to examine efficacy in clinically relevant models. RapidCaP and RAP tissues will be interrogated at molecular and functional levels to examine effects of KBU2046. Impact. Investigators seek to advance the innovative concept that cell motility is an important cancer prevention-interception target. It has applicability to many cancer types.