Evaluation of In Vitro Companion Diagnostic Monoclonal Antibodies for Use in a St

描述（由申请人提供）：本项目旨在允许体外乳腺癌诊断检测开发和/或应用的独立专家Badve博士（乳腺病理学家）和Sledge博士（乳腺肿瘤学家）评价INTICA的候选抗DEspR mAb，以开发商业上可行的伴随诊断检测的初步材料和方法。DEspR是内皮素-1（ET-1）/血管内皮生长因子（VEGF）信号肽（VEGFsp）双重受体，是一种新的、替代的促血管生成/促转移癌症靶标和途径，参与癌症对抗血管生成疗法的耐药性。DEspR在“三阴性”（TNBC;雌激素、孕酮和Her 2受体阴性）乳腺癌（BCa）、胰腺腺癌（PCa）和多形性胶质母细胞瘤（GBM）原发性肿瘤和相应细胞系（MDA-MB-468、Panc-1、U87）中的肿瘤血管内皮细胞（TVEC）、肿瘤细胞（TC）和癌症干细胞（CSC）上表达。INTICA正在开发INTI-1，这是一种抗DEspR治疗性mAb，用于治疗DEspR+癌症，如TNBC、PCa和GBM，尽管TVEC和一些TC上存在VEGF，但抗血管生成治疗无效。体外抗DEspR治疗可预防HUVEC血管生成、TC侵袭和CSC肿瘤球形成，并促进CSC失巢凋亡（细胞凋亡），体内抑制DEspR+自发性肿瘤和CSC异种移植物生长。INTICA还在开发一种伴随诊断（CDx）mAb，作为一种商业化的体外CDx器械，用于对DEspR+肿瘤进行分层，以确定患者对INTI-1的反应。本项目的具体目的是：1）生产候选CDx mAb; 2）通过使用候选CDx mAb的免疫组织化学染色（IHCS）来调查人原发性BCa/TNBC肿瘤和肿瘤微阵列（TMA）的DEspR，以：a）开发IHCS方法，B）将TVEC、TC和CSC上的DEspR表达与肿瘤特征（例如，分级、恶性程度、侵袭性、血管分布），c）将DEspR表达与Oncotype DX复发评分和存活分析相关联，和d）选择优选的CDx mAb; 3）设计初步的CDx IHCS评分系统以将DEspR+肿瘤分层并鉴定可能受益于CDx IHCS的BCa/TNBC癌症患者。在一些实施方案中，本发明的目的在于提供以下方法：（a）从使用INTI-1的抗DEspR疗法中获益（或不获益）;和4）使用这些CDx方法来选择具有不同DEspR表达的人TNBC细胞系，用于异种移植模型中以测试INTI-1。由此产生的CDx测试将用于INTI-1的I期临床试验。关键词。癌症，TNBC，DEspR，INTI-1，伴随诊断，抗血管生成，抗转移，癌症干细胞简要概述。DEspR作为肿瘤干细胞、肿瘤细胞和肿瘤血管的新靶点，以及参与肿瘤转移、复发和血管生成的通路，其发现为肿瘤的治疗提供了新的思路。抗DEspR伴随诊断和治疗mAb的联合使用有可能改变肿瘤学临床实践，特别是在BCa/TNBC中，作为现有治疗的补充或优势。 公共卫生相关性：DEspR作为肿瘤干细胞、肿瘤细胞和肿瘤血管的新靶点，以及参与肿瘤血管生成、转移和复发的通路，其发现为肿瘤治疗提供了新的思路。该项目旨在允许体外乳腺癌诊断测试开发和/或应用的独立专家Badve博士（乳腺病理学家）和Sledge博士（乳腺肿瘤学家）评估INTICA的候选抗DEspR mAb，以开发商业上可行的伴随诊断测试的初步材料和方法。该项目探索了抗DEspR伴随诊断单克隆抗体（mAb）的联合使用，以筛选和分层DEspR+乳腺癌肿瘤，以确定可能受益于（或不受益于）抗DEspR治疗与治疗性mAb INTI-1的患者。正如所设想的，这种方法有可能改变肿瘤学临床实践，特别是在“三阴性”乳腺癌（TNBC）中，作为对现有治疗的补充或优势。

DESCRIPTION (provided by applicant): This project is designed to allow independent experts in the development and/or application of in vitro breast cancer diagnostic tests, Drs. Badve (breast pathologist) and Sledge (breast oncologist), to evaluate INTICA's candidate anti-DEspR mAbs to develop the preliminary materials and methods for a commercially viable companion diagnostic test. DEspR, the dual endothelin-1 (ET-1)/vascular endothelial growth factor (VEGF) signal peptide (VEGFsp) receptor is a novel, alternate pro-angiogenic/pro-metastatic cancer target and pathway involved in cancer resistance to anti-angiogenesis therapies. DEspR is expressed on tumor vascular endothelial cells (TVECs), tumor cells (TCs) and cancer stem cells (CSCs) in "triple-negative" (TNBC; estrogen, progesterone and Her2 receptor-negative) breast cancer (BCa), pancreatic adenocarcinoma (PCa) and glioblastoma multiforme (GBM) primary tumors and respective cell lines (MDA- MB-468, Panc-1, U87). INTICA is developing INTI-1, an Anti-DEspR Therapeutic mAb for use against DEspR+ cancers, such as TNBC, PCa and GBM, where anti-angiogenic therapies are ineffective despite the presence of VEGFRs on TVECs and some TCs. Anti-DEspR therapy in vitro prevents HUVEC angiogenesis, TC invasiveness and CSC tumorsphere formation and promotes CSC anoikis (apoptosis), and in vivo inhibits DEspR+ spontaneous tumor and CSC xenograft growth. INTICA is also developing a Companion Diagnostic (CDx) mAb as a commercial in vitro CDx device to stratify DEspR+ tumors for patient response to INTI-1. The Specific Aims of this project are: 1) to manufacture candidate CDx mAbs; 2) to survey human primary BCa/TNBC tumors and tumor microarrays (TMAs) for DEspR by immunohistochemical staining (IHCS) with candidate CDx mAbs, to: a) develop IHCS methods, b) correlate DEspR expression on TVECs, TCs and CSCs with tumor characteristics (e.g., grade, malignancy, invasiveness, vascularity), c) correlate DEspR expression with Oncotype DX" recurrence scores and survival analysis, and d) select the preferred CDx mAb; 3) to design a preliminary CDx IHCS scoring system to stratify DEspR+ tumors and identify BCa/TNBC cancer patients likely to benefit (or not benefit) from anti- DEspR therapy with INTI-1; and 4) to use these CDx methods to select human TNBC cell lines with varying DEspR expression for use in xenograft models to test INTI-1. The resulting CDx test will be used in Phase 1 clinical trials of INTI-1. Key Words. Cancer, TNBC, DEspR, INTI-1, companion diagnostic, anti-angiogenic, anti-metastatic, cancer stem cells Brief Summary. The discovery of DEspR, a novel target on cancer stem cells, tumor cells and tumor blood vessels, and pathway involved in cancer metastasis, recurrence and angiogenesis, provides a new treatment paradigm. Combined use of anti-DEspR companion diagnostic and therapeutic mAbs has the potential to alter oncology clinical practice, particularly in BCa/TNBC, as an addition to or advantage over existing treatments. PUBLIC HEALTH RELEVANCE: The discovery of DEspR, a novel target on cancer stem cells, tumor cells and tumor blood vessels, and a pathway involved in cancer angiogenesis, metastasis and recurrence provides a new cancer treatment paradigm. This project is designed to allow independent experts in the development and/or application of in vitro breast cancer diagnostic tests, Drs. Badve (breast pathologist) and Sledge (breast oncologist), to evaluate INTICA's candidate anti-DEspR mAbs to develop the preliminary materials and methods for a commercially viable companion diagnostic test. This project explores the combined use of an anti-DEspR companion diagnostic monoclonal antibody (mAb) to screen and stratify DEspR+ breast cancer tumors to identify patients likely to benefit (or not benefit) from anti-DEspR therapy with a therapeutic mAb, INTI-1. As envisioned, this approach has the potential to alter oncology clinical practice, particularly in "triple-negative" breast cancer (TNBC), as an addition to or advantage over existing treatments.