Pianetti Stefania, Miller Kathy D, Chen Hannah H, Althouse Sandra, Cao Sha, Michael Steven J, Sonenshein Gail E, Mineva Nora D
Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA, 02111, USA.
Adecto Pharmaceuticals, Inc., 75 Kneeland St., 14th Floor, Boston, MA, 02111, USA.
Cancer Cell Int. 2023 Aug 11;23(1):165. doi: 10.1186/s12935-023-03024-3.
Breast malignancies are the predominant cancer-related cause of death in women. New methods of diagnosis, prognosis and treatment are necessary. Previously, we identified the breast cancer cell surface protein ADAM8 as a marker of poor survival, and a driver of Triple-Negative Breast Cancer (TNBC) growth and spread. Immunohistochemistry (IHC) with a research-only anti-ADAM8 antibody revealed 34.0% of TNBCs (17/50) expressed ADAM8. To identify those patients who could benefit from future ADAM8-based interventions, new clinical tests are needed. Here, we report on the preclinical development of a highly specific IHC assay for detection of ADAM8-positive breast tumors.
Formalin-fixed paraffin-embedded sections of ADAM8-positive breast cell lines and patient-derived xenograft tumors were used in IHC to identify a lead antibody, appropriate staining conditions and controls. Patient breast cancer samples (n = 490) were used to validate the assay. Cox proportional hazards models assessed association between survival and ADAM8 expression.
ADAM8 staining conditions were optimized, a lead anti-human ADAM8 monoclonal IHC antibody (ADP2) identified, and a breast staining/scoring control cell line microarray (CCM) generated expressing a range of ADAM8 levels. Assay specificity, reproducibility, and appropriateness of the CCM for scoring tumor samples were demonstrated. Consistent with earlier findings, 36.1% (22/61) of patient TNBCs expressed ADAM8. Overall, 33.9% (166/490) of the breast cancer population was ADAM8-positive, including Hormone Receptor (HR) and Human Epidermal Growth Factor Receptor-2 (HER2) positive cancers, which were tested for the first time. For the most prevalent HR-positive/HER2-negative subtype, high ADAM8 expression identified patients at risk of poor survival.
Our studies show ADAM8 is widely expressed in breast cancer and provide support for both a diagnostic and prognostic value of the ADP2 IHC assay. As ADAM8 has been implicated in multiple solid malignancies, continued development of this assay may have broad impact on cancer management.
乳腺恶性肿瘤是女性癌症相关死亡的主要原因。需要新的诊断、预后和治疗方法。此前，我们将乳腺癌细胞表面蛋白ADAM8鉴定为生存不良的标志物，以及三阴性乳腺癌（TNBC）生长和扩散的驱动因素。使用仅用于研究的抗ADAM8抗体进行免疫组织化学（IHC）检测发现，34.0%的TNBC（17/50）表达ADAM8。为了确定那些可能从未来基于ADAM8的干预措施中受益的患者，需要新的临床试验。在此，我们报告一种用于检测ADAM8阳性乳腺肿瘤的高特异性IHC检测方法的临床前开发情况。
使用ADAM8阳性乳腺细胞系和患者来源的异种移植肿瘤的福尔马林固定石蜡包埋切片进行IHC检测，以确定主要抗体、合适的染色条件和对照。使用患者乳腺癌样本（n = 490）对该检测方法进行验证。Cox比例风险模型评估生存与ADAM8表达之间的关联。
优化了ADAM8染色条件，鉴定出一种主要的抗人ADAM8单克隆IHC抗体（ADP2），并构建了一个表达一系列ADAM8水平的乳腺染色/评分对照细胞系微阵列（CCM）。证明了该检测方法的特异性、可重复性以及CCM对肿瘤样本评分的适用性。与早期研究结果一致，36.1%（22/61）的患者TNBC表达ADAM8。总体而言，33.9%（166/490）的乳腺癌患者为ADAM8阳性，包括首次检测的激素受体（HR）和人表皮生长因子受体2（HER2）阳性癌症。对于最常见的HR阳性/HER2阴性亚型，高ADAM8表达可识别出生存不良风险的患者。
我们的研究表明ADAM8在乳腺癌中广泛表达，并为ADP2 IHC检测方法的诊断和预后价值提供了支持。由于ADAM8与多种实体恶性肿瘤有关，该检测方法的持续开发可能对癌症管理产生广泛影响。
