Targeting Cell Cycle Plasticity in Pancreatic Ductal Adenocarcinoma

靶向胰腺导管腺癌的细胞周期可塑性

• 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
• 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

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. !

目前迫切需要开发胰腺导管腺癌的靶向治疗方案 （PDAC），这仍然是一种治疗难治性疾病。数据表明，两个特定的扰动 K-Ras和CDKN 2A基因在PDAC中几乎是通用的。基于这种突变谱，PDAC是 最初被认为是CDK 4抑制疗法的良好候选者。然而，大多数PDAC细胞系和PDX 模型是耐药的，这似乎是由于另一种激酶CDK 2的激活。CDK 4/6特异性 CDK 4 i抑制剂被批准用于转移性HR+乳腺癌。然而，虽然CDK 4 i治疗最初 增加ER+转移性乳腺癌（BC）患者的无进展生存期，最终大多数 由于CDK 2的补偿，产生继发性抗性。因此，CDK 4 i难治性乳腺癌和 PDAC也通过相同的机制出现抗性。因此，为了产生更好的转移性乳腺癌抑制剂， 对于癌症患者和PDAC，需要抑制驱动癌症的激酶（CDK 4/6）和激酶 引起抗性（CDK 2）。CDK 2小分子抑制剂还靶向高度保守和必需的CDK 2受体。 CDK 1激酶，在临床上遇到了不可接受的毒性，因此这种方法已被证明是不成功的。 在这个探索性的R21应用中，我们提出了一种新的策略。而不是针对保守的 CDK 4/6和CDK 2的活性位点，我们靶向p27 Kip 1，它是CDK组装因子，CDK ON/OFF 开关.酪氨酸激酶乳腺肿瘤相关激酶（Brk）对p27（pY 88）的酪氨酸（Y）磷酸化 引起与CDK结合的p27的构象变化，使它们“打开”。我们证明了在乳房中， 用治疗肽ALT阻断p27磷酸化的癌细胞灭活CDK 4/6和CDK 2 为阻断ER+ BC细胞增殖、诱导衰老并最终导致细胞凋亡提供了新的途径 死亡，这在BC动物模型中转化为肿瘤消退和OS增加。我们已经证明 p27在几种已建立的PDAC细胞系中被Y磷酸化，并且在两种细胞系中ALT表达 抑制增殖，表明CDK 2可能被ALT介导的PDAC中p27靶向抑制。 我们的假设是治疗肽ALT抑制CDK 4/6和CDK 2将阻断细胞增殖。 PDAC代表了一种靶向这种肿瘤类型的新方法。我们的具体目标：1）检查效果 ALT和pY阻断疗法对PDAC细胞系的体外作用。2)检验治疗效果 PDAC小鼠模型中的pY阻断疗法和体内ALT。一组原代患者来源的细胞 (PDC)已经测序和表征的品系，以及交叉匹配的PDX模型将用于 测试ALT对细胞生长和肿瘤进展的影响。ALT将通过两种模式输送：诱导型 表达和新的脂质体-肽制剂。这个探索性的R21项目将测试假设， 在临床相关的体外和体内模型中靶向p27，作为概念验证方法，可能代表了 可行的临床策略，应进一步探讨。 !