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

描述(由申请人提供)：该项目旨在允许体外乳腺癌诊断测试开发和/或应用方面的独立专家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单抗，以开发商业上可行的配套诊断测试的初步材料和方法。Despr是一种双重内皮素-1(ET-1)/血管内皮生长因子(VEGF)信号肽(VEGFsp)受体，是一种新的、交替的促血管生成/促转移的肿瘤靶点和途径，参与肿瘤对抗血管生成治疗的抵抗。DespR在三阴性乳腺癌(BCA)、胰腺癌(PCa)和多形性胶质母细胞瘤(GBM)原发肿瘤及相应细胞系(MDA-MB-468、PANC-1、U87)的肿瘤血管内皮细胞(TVECs)、肿瘤细胞(TCS)和肿瘤干细胞(CSCs)上均有表达。INTICA正在开发Inti-1，一种用于治疗Despr+癌症的抗Despr治疗单抗，如TNBC、PCa和GBM，在这些癌症中，尽管TVEC和一些TC上存在VEGFR，但抗血管生成治疗无效。体外抗Despr治疗可阻止HUVEC血管生成、TC侵袭和CSC肿瘤球体的形成，促进CSC的凋亡，在体内抑制Despr+自发性肿瘤和CSC异种移植瘤的生长。INTICA还在开发一种伴生诊断(CDX)单抗，作为一种商业体外CDX设备，用于对Despr+肿瘤进行分层，以评估患者对Inti-1的反应。本项目的具体目标是：1)制备候选CDX单抗；2)用候选CDX单抗通过免疫组织化学染色(IHCS)检测人BCA/TNBC肿瘤和肿瘤微阵列(TMA)中Despr的表达，a)建立IHCS方法，b)将Despr在TVECs、TCS和CSCs上的表达与肿瘤特征(如分级、恶性程度、侵袭性、血管密度)相关联，c)将Despr表达与Oncotype DX复发评分和生存分析相关联，以及d)选择首选的CDX单抗；3)设计一个初步的CDX IHCS评分系统来对Despr+肿瘤进行分层，并确定可能受益于(或不受益)Inti-1抗Despr治疗的BCA/TNBC癌症患者；以及4)使用这些CDX方法选择Despr表达不同的人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的第一阶段临床试验。关键词。癌症，TNBC，Despr，Inti-1，伴随诊断，抗血管生成，抗转移，癌症干细胞简介。Despr是肿瘤干细胞、肿瘤细胞和肿瘤血管的新靶点，是肿瘤转移、复发和血管生成的重要途径，它的发现为肿瘤的治疗提供了新的思路。联合使用抗Despr联合诊断和治疗单抗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.