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

描述（由申请人提供）：本项目旨在允许独立专家在开发和/或应用体外乳腺癌诊断测试，博士。Badve（乳腺病理学家）和Sledge（乳腺肿瘤学家），评估INTICA的候选抗despr单克隆抗体，为商业上可行的伴随诊断测试开发初步材料和方法。desr是内皮素-1 (ET-1)/血管内皮生长因子（VEGF）信号肽（VEGFsp）的双重受体，是一种新的促血管生成/促转移的癌症靶点和途径，参与癌症对抗血管生成治疗的抵抗。在“三阴性”（TNBC；雌激素、孕激素和Her2受体阴性）乳腺癌（BCa）、胰腺腺癌（PCa）和多形性胶质母细胞瘤（GBM）原发性肿瘤及其细胞系（MDA- MB-468、Panc-1、U87）中，DEspR表达于肿瘤血管内皮细胞（TVECs）、肿瘤细胞（TCs）和癌症干细胞（CSCs）上。INTICA正在开发一种抗desr治疗单抗，用于治疗desr +癌症，如TNBC、PCa和GBM，尽管在tvec和一些tc上存在vegfr，但抗血管生成治疗无效。体外抗DEspR治疗可抑制HUVEC血管生成、TC侵袭和CSC肿瘤球形成，促进CSC细胞凋亡，体内抗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）单抗，作为一种商业化的体外CDx设备，用于对DEspR+肿瘤进行分层，以观察患者对INTI-1的反应。该项目的具体目标是：1)制备候选CDx单克隆抗体；2)用候选CDx单克隆抗体免疫组化染色（IHCS）研究人原发性BCa/TNBC肿瘤和肿瘤微阵列（TMAs）对DEspR的检测，以：a)建立IHCS方法，b)将tvec、tc和CSCs上的DEspR表达与肿瘤特征（如分级、恶性、侵袭性、血管性）联系起来，c)将DEspR表达与Oncotype DX的复发评分和生存分析联系起来，d)选择首选CDx单克隆抗体；3)设计一个初步的CDx IHCS评分系统，对DEspR+肿瘤进行分层，并确定BCa/TNBC癌症患者可能受益（或不受益）抗DEspR治疗与INTI-1；4)使用这些CDx方法选择具有不同DEspR表达的人类TNBC细胞系用于异种移植模型以检测INTI-1。由此产生的CDx测试将用于INTI-1的1期临床试验。关键字。癌症，TNBC, DEspR, INTI-1，伴随诊断，抗血管生成，抗转移，癌症干细胞。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，DEspR的发现提供了一种新的治疗范式。联合使用抗despr伴随诊断和治疗的单克隆抗体有可能改变肿瘤临床实践，特别是在BCa/TNBC中，作为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.