Development and Preclinical Applications of Pancreatic Adenocarcinoma Models

胰腺癌模型的建立及临床前应用

• 批准号: 9780037
• 负责人: SHYAM SHARAN
• 金额: $ 158.82万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9780037
• 关键词: Adenocarcinoma Cell; Adult; Affect; Alleles; Allografting; Animals; Antineoplastic Agents; Biological; Biological Availability; CCR; Cancer Model; Cells; Characteristics; Chemicals; Clinical; Collaborations; Combined Modality Therapy; Communities; Complex; Computer Simulation; Cultured Cells; Cytotoxic T-Lymphocytes; Data; Data Reporting; Desmoplastic; Development; Diagnosis; Disease; Dose; Drug Targeting; Engineering; Epithelial; Epithelial Cells; Evaluation; Event; Formulation; Gene Expression Profile; Generations; Genetic; Genetic Recombination; Genetically Engineered Mouse; Genomics; Goals; Growth; Human; Immune Targeting; Immune system; Immunocompetent; Immunologics; Immunotherapeutic agent; Immunotoxins; In Vitro; Inflammatory; Interleukin-15; Intervention; Intraepithelial Neoplasia; Investigation; Joints; KRAS2 gene; Laboratories; Lesion; Liver; Liver Circulation; Lung; Malignant Neoplasms; Malignant neoplasm of pancreas; Manuscripts; Mediating; Metastatic to; Minor; Missense Mutation; Modeling; Molecular; Mus; Mutagenesis; Mutation; Neoplasm Metastasis; Nucleotides; Oncogenic; Orthologous Gene; PDCD1LG1 gene; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Pancreatic duct; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phenotype; Pre-Clinical Model; Premalignant; Preparation; Prevalence; Primary Neoplasm; Protein Isoforms; Protein p53; Proteins; Publications; Recombinants; Regimen; Reporting; Research Personnel; Resources; Scientist; Series; Signal Transduction; Subcellular structure; Survival Rate; TP53 gene; Therapeutic; Thyroid Gland; Tissues; Toxic effect; Transcriptional Regulation; Transgenes; Transplantation; Tumor Tissue; Tumor-Derived; Up-Regulation; Variant; anti-PD-1; biomarker discovery; carcinogenicity; cohort; cytokine; design; drug development; efficacy evaluation; experimental study; gemcitabine; immune checkpoint blockade; immunogenic; immunological intervention; improved; in vivo; irradiation; macrophage; mesothelin; mouse model; mutant; neoantigens; next generation; novel; novel strategies; novel therapeutics; pancreatic neoplasm; polypeptide; pre-clinical; pre-clinical research; preclinical efficacy; precursor cell; programs; prototype; rare cancer; receptor; screening; standard of care; therapeutic candidate; tool; transplant model; treatment response; tumor

Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest and clinically burdensome malignancies with a five-year survival rate less than 5%. PDACs are notoriously aggressive, prone to rapid conversion to metastatic phenotype, have remarkable desmoplastic features with strong prevalence of stromal components hindering therapeutic access, and are moreover difficult to diagnose at early stages. Thus, we are executing studies to meet the stated objectives to develop, utilize, and provide to the community several tractable preclinical PDAC models for therapeutic and biomarker discovery. CAPR recent progress on internal initiatives includes generation and full characterization of three relevant Cre-inducible missense p53 GEM strains in which the non-recombined allele expresses wild type p53 prior to a Cre-driven conversion to a mutant isoform expressing one. The p53aa172 inducible mutant strain has been bred with PDX-Cre and KRASG12D-lsl mice to generate animals that are similar to the KPC model, with the exception that they harbor no null allele prior to expression of p53-172. Development of PDA in these mice (KPwt-172C) is similar to that of KPC mice, except that ectopic tumors do not develop and PDA develops with shortened latency. Furthermore, orthotopic KPC-PDA GDA models have been established from several KPC-derived primary tumors and shown to develop PDA with characteristics of the parental malignancy. In addition, the metastatic rate is increased to about 70 %, thus potentially enabling studies on mechanisms and treatment of metastatic disease. This high metastatic rate makes this model ideal for evaluating therapies for metastatic disease. As one of the most visible preclinical resource for intramural CCR investigators, CAPR pancreatic program is involved in multiple collaborative initiatives with CCR PI labs. For example, the pancreatic team has recently finalized a large, multi-modular collaborative project with Dr. Udo Rudloff's lab pursuing a comprehensive IND-enabling characterization of metarrestin compound employing both de novo and syngeneic orthotopic transplant models of pancreatic adenocarcinoma (PDAC). Ten integrated preclinical assessment studies interrogating bioavalability, in vivo toxicity, a spectrum of molecular responses to treatment, as well as efficacy as monotherapy and in combination with gemcitabine standard of care, have been completed and will be used to support the upcoming IND submission for metarrestin. The results of this collaborative effort are being written up as two publications co-authored by CAPR scientists. Expanding collaborative relations with Dr. Rudloff's section of the Rare Tumor initiative, CAPR also made significant progress with the second, CAPR Oversight Committee-approved joint study to evaluate another molecule - a biosimilar peptide compound RP-182 - which has shown promise as a potential microenvironment re-modelling factor in PDAC. CAPR has conducted several pilot multi-organ PK studies to explore bioavailability in pancreatic tumors, other PDAC-affected tissues (e.g. lung and liver) and circulation. CAPR has also conducted preclinical efficacy evaluation experiments to assess the anti-tumor potency of RP-182 peptide in KPC model, as well as in orthotopic pancreatic cancer models established using wild-type and CD206-deficient syngeneic recipient animals to confirm the hypothesis of CD206 receptor being a specific target molecule for RP-182 peptide. Another CCR PI-initiated collaborative PDAC study is ongoing with Dr. Martin Schnermann's lab. In this project, CAPR joined efforts with drug development chemists to assess a novel strategy of attacking this lethal malignancy with photo-activated formulations synthesized to selectively release an anti-cancer drug payload in disease-affected tissues upon near-IR irradiation. CAPR is currently assisting Dr. Schnermann's team in optimizing chemical composition and delivery mechanisms for photo-activated therapeutic candidate compounds in KPC pancreatic models. CAPR has also completed a collaborative study with Dr. William Stetler-Stevenson's lab to assess the ability of TIMP-2 recombinant polypeptide to suppress the cancer growth and metastatic dissemination of PDAC in the autochthonous KPC mouse model. Currently, the project is finalized and the study report is being prepared. A joint project with Dr. Peter Johnson's lab is underway to investigate the biological significance of Perinuclear Signaling Complexes (PSC) characteristic for Ras-driven malignancies in carcinogenic progression, and to validate these intriguing intracellular structures as possible drug targets to ameliorate aggressive cancers featuring deregulated Ras pathways. CAPR pancreatic program scientists also completed a collaborative project with Dr. George Pavlakis' laboratory aimed at investigation of immunologic signatures in pancreatic tumors treated with recombinant hetIL-15 cytokine. Pancreatic tumor bearing animals have been subject to dosing with control vehicle compound or with IL-15 and gemcitabine as single drug regimens, or IL-15/Gemcitabine combination therapy. CAPR is currently evaluating the data from this study. Continuing to deliver on its collaboration with Dr. Christine Alewine, CAPR researchers established a broad allelic series of genetically modified mouse models expressing either chimeric mouse/human mesothelin (MSLN) protein under the transcriptional control of the endogenous mMSLN locus, or a fully human MSLN ortholog in a narrow expression domain in the thyroid gland. Having undergone extensive molecular characterization, these validated new tools provide immunocompetent recipient animals tolarized against hMSLN protein or lacking MSLN expression in either a constitutive or conditional manner, and will now be used for syngeneic allografting studies with PDAC cells expressing chimMSLN protein to prepare cohorts of tumor bearing animals to assess the efficacy of recombinant immunotoxins designed to recognize human MSLN protein. Two manuscripts reporting data from these collaborative efforts are in advanced stage of preparation.

胰腺导管腺癌 (PDAC) 是最致命且临床负担最重的恶性肿瘤之一，五年生存率低于 5%。 PDAC 具有众所周知的侵袭性，易于快速转化为转移表型，具有显着的促纤维增生特征，基质成分的普遍存在阻碍了治疗的获得，而且在早期阶段难以诊断。因此，我们正在进行研究，以实现既定目标，开发、利用并向社区提供几种易于处理的临床前 PDAC 模型，用于治疗和生物标志物发现。 CAPR 最近在内部计划方面取得的进展包括三种相关 Cre 诱导错义 p53 GEM 菌株的生成和完整表征，其中非重组等位基因表达野生型 p53，然后再由 Cre 驱动转换为表达野生型 p53 的突变同工型。 p53aa172 诱导突变株已与 PDX-Cre 和 KRASG12D-lsl 小鼠一起培育，产生与 KPC 模型相似的动物，不同之处在于它们在表达 p53-172 之前不存在无效等位基因。这些小鼠 (KPwt-172C) 中 PDA 的发育与 KPC 小鼠相似，只是异位肿瘤不发育并且 PDA 发育潜伏期缩短。此外，已经从几个 KPC 衍生的原发性肿瘤中建立了原位 KPC-PDA GDA 模型，并显示出可发展具有亲代恶性肿瘤特征的 PDA。此外，转移率增加至约70%，因此有可能使研究转移性疾病的机制和治疗成为可能。这种高转移率使该模型成为评估转移性疾病治疗的理想选择。作为壁内 CCR 研究人员最明显的临床前资源之一，CAPR 胰腺项目参与了与 CCR PI 实验室的多项合作计划。例如，胰腺团队最近与 Udo Rudloff 博士的实验室完成了一个大型、多模块合作项目，旨在利用胰腺腺癌 (PDAC) 的从头和同基因原位移植模型对metarrestin 化合物进行全面的 IND 表征。十项综合临床前评估研究已经完成，涉及生物利用度、体内毒性、治疗的一系列分子反应以及单一疗法和与吉西他滨护理标准相结合的疗效，这些研究已完成，并将用于支持即将提交的metarrestin IND 申请。这项合作的成果将由 CAPR 科学家共同撰写的两本出版物撰写。 CAPR 扩大了与 Rudloff 博士罕见肿瘤计划部门的合作关系，并在 CAPR 监督委员会批准的第二项联合研究中取得了重大进展，该研究评估了另一种分子——生物类似肽化合物 RP-182——该分子已显示出作为潜在药物的前景。 PDAC 中的微环境重塑因子。 CAPR 进行了多项试点多器官 PK 研究，以探索胰腺肿瘤、其他 PDAC 影响的组织（例如肺和肝）和循环中的生物利用度。 CAPR还进行了临床前疗效评估实验，以评估RP-182肽在KPC模型以及使用野生型和CD206缺陷同系受体动物建立的原位胰腺癌模型中的抗肿瘤效力，以证实CD206的假设受体是 RP-182 肽的特异性靶分子。另一项由 CCR PI 发起的 PDAC 合作研究正在与 Martin Schnermann 博士的实验室进行。在该项目中，CAPR 与药物开发化学家共同努力，评估一种利用光激活制剂攻击这种致命恶性肿瘤的新策略，该制剂合成后可在近红外照射下选择性地在受疾病影响的组织中释放抗癌药物有效负载。 CAPR 目前正在协助 Schnermann 博士的团队优化 KPC 胰腺模型中光激活治疗候选化合物的化学成分和递送机制。 CAPR 还与 William Stetler-Stevenson 博士的实验室完成了一项合作研究，以评估 TIMP-2 重组多肽在本地 KPC 小鼠模型中抑制 PDAC 癌症生长和转移性传播的能力。目前，该项目已定稿，正在编写研究报告。与 Peter Johnson 博士实验室的联合项目正在进行中，旨在研究核周信号复合物 (PSC) 特征对于 Ras 驱动的恶性肿瘤在致癌进展中的生物学意义，并验证这些有趣的细胞内结构作为可能的药物靶点，以改善以失调为特征的侵袭性癌症Ras 途径。 CAPR 胰腺项目的科学家还与 George Pavlakis 博士的实验室完成了一个合作项目，旨在研究重组 hetIL-15 细胞因子治疗的胰腺肿瘤的免疫特征。患有胰腺肿瘤的动物已经接受了对照媒介物化合物或IL-15和吉西他滨作为单一药物方案的给药，或IL-15/吉西他滨联合治疗。 CAPR 目前正在评估这项研究的数据。 CAPR 研究人员继续与 Christine Alewine 博士合作，建立了一系列广泛的等位基因转基因小鼠模型，这些模型在内源性 mMSLN 基因座的转录控制下表达嵌合小鼠/人间皮素 (MSLN) 蛋白，或表达完全人 MSLN甲状腺中狭窄表达域的直向同源物。经过广泛的分子表征后，这些经过验证的新工具以组成型或条件型方式提供了针对 hMSLN 蛋白或缺乏 MSLN 表达的免疫活性受体动物，现在将用于表达 chimMSLN 蛋白的 PDAC 细胞的同基因同种异体移植研究，以制备肿瘤组生育动物评估旨在识别人类 MSLN 蛋白的重组免疫毒素的功效。报告这些合作成果数据的两份手稿正处于后期准备阶段。