Antibody targeting of ADAM8 for treatment of triple-negative breast cancer

三阴性乳腺癌(TNBCs)约占乳腺癌死亡的25%，缺乏针对性的治疗。 Sonenshein、Mineva和Romagnoli博士以及他们的同事最近发现了非必需的细胞表面 蛋白ADAM8(A去整合素和金属蛋白酶8)作为乳腺肿瘤生长和生长的关键促进剂 转移，并验证其作为TNBC抗体治疗的靶点。ADAM8基因在卵巢癌组织中高表达 TNBCs，其水平与患者预后不良相关。ADAM8蛋白在34%的原发肿瘤中存在 TNBCs和一半的乳腺癌患者起源的转移，但在邻近的正常乳腺组织中没有。 来自ADAM8基因敲除的TNBC细胞的原位肿瘤只生长到可以触摸到的大小，并且产生的肿瘤非常少 循环中的肿瘤细胞和脑转移。金属蛋白酶(MP)和去整合素(DI)结构域 ADAM8在肿瘤生长和扩散中起关键作用，它通过释放促血管生成因子和 分别激活β-1整合素对癌细胞的作用。试剂级商品抗ADAM8的治疗 小鼠单抗MAB1031(研发)，对MP和Mp均有体外拮抗活性 Di结构域，1.5 mg/kg时，与对照同型相合的IgG2B相比，原发TNBC肿瘤负担降低70% 在细胞植入时开始的原位模型。MAB1031也大大减少了 先前存在的肿瘤扩散到脑和肺，提供了双重拮抗剂的概念证明 可以制备单抗，并且这两个区域都可以在体内进行基于抗体的治疗。因此，我们 假设基于ADAM8抗体的治疗构成了对TNBC患者的有效治疗 表达这种跨膜蛋白。PCT专利申请PCT/US14/37857于2014年5月13日由 Sonenshein博士、Mineva博士和Romagnoli博士以及塔夫茨大学的指控包括以ADAM8为目标 MP和DI结构域用于治疗乳腺癌和其他ADAM8驱动的癌症，包括胰腺癌 腺癌。2014年10月，Adecto PharmPharmticals，Inc.(AP)由三位发明家与 开发ADAM8特异性抗体治疗TNBC和转移性乳腺癌的目标 最初的迹象是。在此阶段1 STTR应用中，AP将与Sonenshein实验室(SL)密切合作，以 制备同时抑制其MP和DI结构域的人ADAM8特异性鼠单抗并进行中试 在小鼠身上进行临床前试验。其具体目的是：(1)分离一组针对带有MP的HuADAM8的单抗 和DI域拮抗剂活性；(2)用细胞测定法确定最有效的拮抗剂单抗；(3) 对两种最有效的ADAM8拮抗剂单抗抑制生长的能力进行体内中试测试 以及先前存在的荧光素酶标记的MDA-MB-231细胞来源的肿瘤的转移性扩散。我们建议 基于ADAM8抗体的治疗有可能使TNBC患者的治疗发生革命性变化，以及 降低与转移性乳腺癌相关的死亡率，因此将成为护理的新组成部分 为TNBC工作。

Triple-negative breast cancers (TNBCs) account for ~25% of breast cancer deaths and lack targeted therapies. Drs. Sonenshein, Mineva and Romagnoli and their co-workers recently identified the non-essential, cell surface protein ADAM8 (A Disintegrin And Metalloprotease 8) as a pivotal promoter of breast tumor growth and metastasis, and validated it as a target of antibody therapy for TNBC. ADAM8 mRNA was highly expressed in TNBCs, and its level correlated with poor patient outcome. ADAM8 protein was present in 34% of primary TNBCs, and half of all breast cancer patient-derived metastases, but absent in adjacent normal breast tissues. Orthotopic tumors from ADAM8 knockdown TNBC cells grew only to a palpable size and generated very few circulating tumor cells and brain metastases. The Metalloproteinase (MP) and Disintegrin (DI) domains of ADAM8 were critical in tumor growth and dissemination through release of pro-angiogenic factors and activation of β1-integrin on cancer cells, respectively. Treatment with a reagent grade commercial anti-ADAM8 mouse monoclonal antibody (mAb) MAB1031 (R&D), with in vitro antagonist activity against both the MP and DI domains, reduced primary TNBC tumor burden by 70% at 1.5 mg/kg vs control isotype-matched IgG2B in an orthotopic model when started at the time of cell implantation. MAB1031 also profoundly reduced dissemination of pre-existing tumors to the brain and lungs, providing proof-of-concept that a dual antagonist mAb can be prepared and that both domains are accessible to antibody-based therapy in vivo. Thus, we hypothesize that ADAM8 antibody-based treatment constitutes an effective therapy for TNBC patients expressing this transmembrane protein. A PCT patent application PCT/US14/37857 was filed May 13, 2014 by Drs. Sonenshein, Mineva and Romagnoli, and Tufts University, with claims including the targeting of ADAM8 MP and DI domains for treatment of breast and other ADAM8-driven cancers, including pancreatic adenocarcinomas. In October 2014, Adecto Pharmaceuticals, Inc (AP) was founded by the three inventors with the goal of developing ADAM8-specific antibodies for the treatment of TNBC and metastatic breast cancer as the initial indications. In this Phase 1 STTR application, AP will work closely with the Sonenshein lab (SL) to prepare mouse mAbs specific for human ADAM8 that inhibit both its MP and DI domains and perform pilot preclinical testing in mice. The specific aims are to: (1) Isolate a panel of mAbs specific for HuADAM8 with MP and DI domain antagonist activity; (2) Identify the most effective antagonist mAbs using cell based assays; (3) Perform pilot in vivo testing of the ability of the two most effective ADAM8 antagonist mAbs to inhibit growth and metastatic dissemination of pre-existing luciferase-tagged MDA-MB-231 cell-derived tumors. We propose that ADAM8 antibody-based therapy has the potential to revolutionize the treatment of TNBC patients, and reduce the mortality associated with metastatic breast cancer and thus will become a new component of care for TNBC.