Targeting Cell Cycle Plasticity in Pancreatic Ductal Adenocarcinoma

靶向胰腺导管腺癌的细胞周期可塑性

基本信息

批准号：
10044497
负责人：
Chongmin Huan
金额：
$ 43.83万
依托单位：
SUNY DOWNSTATE MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10044497
关键词：
Active Sites Animal Cancer Model Animals Apoptosis Automobile Driving Breast Cancer Cell Breast Cancer Model Breast Cancer Patient Breast Cancer cell line CDC2 gene CDK2 gene CDK4 gene CDKN2A gene Cell Cycle Cell Cycle Proteins Cell Death Cell Line Cell Proliferation Cells Clinic Clinical Cyclin D1 Data Dependence Development Disease Doxycycline Drug Delivery Systems Early Diagnosis Engineering Epigenetic Process Estrogen receptor positive Financial compensation Formulation Genes Genetic Goals In Vitro Injections KRAS2 gene Lead Liposomes Malignant Neoplasms Mammary Neoplasms Mediating Metastatic breast cancer Modality Modeling Molecular Conformation Mutation Necrosis Operative Surgical Procedures Oral Pancreatic Ductal Adenocarcinoma Patient-Focused Outcomes Patients Peptides Phosphorylation Phosphotransferases Progression-Free Survivals Protein Tyrosine Kinase Refractory Resistance Signal Pathway Signal Transduction Survival Rate Tail Testing Therapeutic Effect Toxic effect Translating Tumor Volume Tyrosine Veins Work Xenograft Model base cell growth clinically relevant in vivo in vivo Model inducible gene expression inhibitor/antagonist malignant breast neoplasm mouse model mutational status neoplastic cell novel novel strategies peptide drug prevent protein expression response senescence small molecule inhibitor targeted treatment therapeutic target transcriptome tumor tumor progression

项目摘要

There is an urgent unmet need to develop targeted therapeutic options for Pancreatic Ductal Adenocarcinoma (PDAC), which continues to be a therapy recalcitrant disease. Data has shown that perturbation of two specific genes, K-Ras and CDKN2A, are nearly universal in PDAC. Based on this mutational profile, PDAC was thought initially to be a good candidate for CDK4 inhibition therapy. However, most PDAC cell lines and PDX models are resistant and this appears to be due to the activation of another kinase CDK2. The CDK4/6 specific inhibitors (CDK4i) are approved for metastatic HR+ breast cancer. However, while CDK4i therapy initially increases progression-free survival in ER+ metastatic breast cancer (BC) patients, ultimately the majority develop secondary resistance, due to compensation by CDK2. Thus, both CDK4i-refractory breast cancer and PDAC appear resistant by the same mechanism. Therefore, to generate a better inhibitor for metastatic breast cancer patients and PDAC, one needs to inhibit both the kinase driving cancer (CDK4/6) and the kinase causing resistance (CDK2). CDK2 small molecule inhibitors also target the highly conserved and essential CDK1 kinase and have met with unacceptable toxicity in the clinic, so this approach has proven unsuccessful. In this exploratory R21 application, we propose using a novel strategy. Instead of targeting the conserved active sites of CDK4/6 and CDK2, we target p27Kip1, which is a CDK assembly factor and CDK ON/OFF switch. Tyrosine (Y) phosphorylation of p27 (pY88) by the tyrosine kinase Breast tumor Related Kinase (Brk) causes a conformational change in p27 bound to CDKs, turning them “ON”. We demonstrated that in breast cancer cells blocking phosphorylation of p27 with the therapeutic peptide ALT inactivated CDK4/6 and CDK2 and represented a new way to block ER+ BC cell proliferation, induce senescence and ultimately cause cell death, which translates into tumor regression and increased OS in BC animal models. We have shown that p27 is Y phosphorylated in several established PDAC cell lines, and expression of ALT in two cell lines inhibited proliferation, suggesting that CDK2 might be inhibited by ALT-mediated targeting of p27 in PDAC. Our hypothesis is that inhibition of CDK4/6 and CDK2 by the therapeutic peptide ALT will block proliferation of PDAC and represents a novel way to target this tumor type. Our specific Aims: 1) To examine the effect that ALT and pY blockage therapy has on PDAC cell lines in vitro. 2) To examine the therapeutic effect pY blockage therapy and ALT in vivo in PDAC mouse models. A panel of primary patient derived Cell (PDC) lines, which have been sequenced and characterized, and cross-matched PDX models will be used to test ALT's effect on cell growth and tumor progression. ALT will be delivered by two modes: inducible expression and a novel liposomal-peptide formulation. This exploratory R21 project will test the hypothesis that targeting p27 in clinically relevant in vitro and in vivo models, as a proof of concept approach, may represent a viable clinical strategy, which should be further explored. !

迫切需要开发针对胰腺导管腺癌的有针对性的治疗选择（PDAC），它仍然是一种治疗顽固性疾病。数据表明，两个特定的扰动基因，K-RAS和CDKN2A在PDAC中几乎是通用的。基于此突变概况，PDAC为最初认为是CDK4抑制疗法的良好候选者。但是，大多数PDAC细胞系和PDX 模型具有抗性，这似乎是由于另一个激酶CDK2的激活。特定于CDK4/6 抑制剂（CDK4I）被批准用于转移性HR+乳腺癌。但是，虽然CDK4I疗法最初提高ER+转移性乳腺癌（BC）患者的无进度生存率，最终大多数由于CDK2的补偿，发育次级电阻。这两个cdk4i-抗乳性乳腺癌和 PDAC通过相同的机制似乎具有抵抗力。因此，为转移性乳房产生更好的抑制剂癌症患者和PDAC，需要抑制激酶驱动癌症（CDK4/6）和激酶引起电阻（CDK2）。 CDK2小分子抑制剂还针对高度保守和必不可少的 CDK1激酶并在诊所遇到了不可接受的毒性，因此这种方法被证明没有成功。在此探索性R21应用中，我们建议使用一种新型策略。而不是针对保守的 CDK4/6和CDK2的活性位点，我们靶向P27KIP1，这是CDK组装因子，CDK ON/OFF 转变。酪氨酸激酶乳腺肿瘤相关激酶（BRK）对p27（PY88）的酪氨酸（Y）磷酸化（PY88）导致p27与CDK的构象变化，并将其“打开”。我们证明了在乳房中用治疗肽Alt灭活CDK4/6和CDK2阻断p27磷酸化的癌细胞并代表了阻断ER+ BC细胞增殖的一种新方法，诱导感应并最终引起细胞死亡，转化为卑诗省动物模型中的肿瘤回归和OS增加。我们已经表明 p27在几种已建立的PDAC细胞系中磷酸化，在两个细胞系中的表达抑制增殖，表明CDK2可能由于P27在PDAC中的p27靶向抑制。我们的假设是，治疗性胡椒对CDK4/6和CDK2的抑制作用将阻止 PDAC代表了靶向这种肿瘤类型的一种新型方法。我们的具体目的：1）检查效果 ALT和PY阻滞疗法在体外对PDAC细胞系具有。 2）检查治疗效果 PY阻滞疗法和PDAC小鼠模型中的体内体内。一组原代患者衍生细胞（PDC）已被测序和表征的线，交叉匹配的PDX模型将用于测试ALT对细胞生长和肿瘤进展的影响。 Alt将通过两种模式传递：可诱导表达和一种新型的脂质体肽式。这个探索性R21项目将检验以下假设针对临床相关的体外和体内模型中的p27作为概念证明，可能代表可行的临床策略，应进一步探讨。呢