Inflammatory Breast Cancer: Factors Contributing to Dissemination

描述(由申请人提供)：IBC是最致命的原发性乳腺癌，主要针对年轻女性。IBC占美国所有乳腺癌病例的5%，但其在非裔美国女性中的发病率明显更高(10.1%)。在埃及，IBC的发病率据报道占所有乳腺癌病例的5%-10%，而且还在上升。广泛的皮肤淋巴管侵袭和肿瘤栓子导致淋巴管阻塞，这是该病炎症性质的基础。我们发现巨噬细胞渗入IBC的微环境，包围IBC栓子，并存在于IBC患者的腋血中。我们在这项建议中的工作假设是，IBC相关巨噬细胞在肿瘤栓子的形成和癌细胞在淋巴管中的扩散中发挥关键作用，这是IBC的两个表型特征。FIRCA建议的具体目标是：1)建立IBC细胞的3D培养和a)评估侵袭性表型；b)确定IBC标记蛋白的表达/定位；c)定量细胞因子、趋化因子和生长因子在条件培养液中的表达；以及d)半胱氨酸组织蛋白酶的表达/分泌/活性；2)表征IBC和非IBC患者的癌栓、肿瘤、阳性淋巴结、腋窝分支血和外周血中单核/巨噬细胞的表型和含量；3)从IBC腋窝支流血和外周血分离单核细胞，a)定量分泌的细胞因子、趋化因子和生长因子的表达，以及b)半胱氨酸组织蛋白的表达/分泌/活性；4)IBC标本石蜡切片免疫染色以检测选定的细胞因子、趋化因子和生长因子的细胞表面受体；以及，5)建立IBC细胞和从IBC腋窝支流血和外周血分离的单核细胞的3D共培养，a)评估侵袭表型；b)确定可能的IBC标记蛋白的表达/定位；c)在条件培养液中定量细胞因子、趋化因子和生长因子的表达；以及d)半胱氨酸组织蛋白的表达/分泌/活性；免疫染色共培养IBC标本中鉴定的细胞表面受体。未来的方向将使用在AIMS 1-5中获得的信息来选择半胱氨酸组织蛋白、细胞因子、趋化因子和生长因子，这些可能是降低IBC侵袭性表型的靶点。IBC在埃及的侵略性表现意味着，迫切需要开发能够立即帮助乳腺癌患者的研究项目。我们预计，我们的结果将有助于描述IBC的病理生物学基础，这是优先考虑治疗干预的新靶点的先决条件。这项研究将主要在埃及开罗大学与莫娜·穆斯塔法·穆罕默德合作进行，作为NIH第号补助金的延伸。R01CA131990，8-01-2008至5-31-2013。 与公共健康相关：这项提案汇集了一个由埃及和美国调查人员组成的多学科团队，共同努力，最终目标是找出可以作为针对IBC的新疗法的可用药途径。IBC是一种高度致命的癌症，在年轻女性中发现的比例更高，在埃及发病率很高。本申请中建议的研究的成功完成将导致经过验证的体外模型系统，该系统可用于临床前研究，筛选新的治疗药物，并可能识别IBC的生物标记物。我们的目的是改善这种毁灭性癌症的诊断和最终的治疗管理。

DESCRIPTION (provided by applicant): IBC is the most lethal form of primary breast cancer and disproportionately targets younger women. IBC accounts for <5% of all breast cancer cases in the United States, but its incidence is significantly higher among African-American women (10.1%). In Egypt, the incidence of IBC is reported to be from 5- 10% of all breast cancer cases and is on the rise. Extensive dermal lymphatic invasion and tumor emboli therein result in lymphatic obstruction that underlies the inflammatory nature of the disease. We have shown that macrophages infiltrate the IBC microenvironment, surround IBC emboli and are present in axillary blood of IBC patients. Our working hypothesis in this proposal is that IBC-associated macrophages play a crucial role in the formation of tumor emboli and dissemination of the cancer cells in the lymphatics, two of the phenotypic hallmarks of IBC. The Specific Aims of this FIRCA proposal are to: 1) Establish 3D cultures of IBC cells and a) assess invasive phenotype; b) determine expression/localization of putative IBC marker proteins; c) quantify expression of cytokines, chemokines and growth factors in conditioned media; and d) expression/secretion/activity of cysteine cathepsins; 2) Characterize monocyte/macrophage phenotype and content in tumor emboli, tumors, positive lymph nodes, axillary tributary blood and peripheral blood from IBC and non-IBC patients; 3) Isolate monocytes from IBC axillary tributary blood and peripheral blood and a) quantify expression of secreted cytokines, chemokines and growth factors, and b) expression/secretion/activity of cysteine cathepsins; 4) Immunostain paraffin blocks of IBC specimens for cell surface receptors of selected cytokines, chemokines and growth factors; and, 5) Establish 3D cocultures of IBC cells and monocytes isolated from IBC axillary tributary blood and peripheral blood and a) assess invasive phenotype; b) determine expression/localization of putative IBC marker proteins; c) quantify expression of cytokines, chemokines and growth factors in conditioned media; and d) expression/secretion/activity of cysteine cathepsins; and, e) immunostain cocultures for cell surface receptors identified in IBC specimens. Future directions will use the information obtained in Aims 1-5 to select cysteine cathepsins, cytokines, chemokines and growth factors that might be targeted to reduce the invasive phenotype of IBC. The aggressive manifestation of IBC in Egypt means that there is a pressing need to develop research programs that can be of immediate assistance to patients suffering from breast cancer. We anticipate that our results will contribute to the characterization of the pathobiological underpinnings of IBC, a prerequisite for prioritizing novel targets for therapeutic intervention. This research will be done primarily in Egypt at Cairo University in collaboration with Mona Mostafa Mohamed, as an extension of NIH Grant No. R01CA131990, 8-01-2008 to 5-31-2013. PUBLIC HEALTH RELEVANCE: This proposal brings together a multi-disciplinary team of Egyptian and American investigators in a collaborative effort with the ultimate goal of identifying druggable pathways that can be targeted for new therapies against IBC, a highly lethal cancer that is found disproportionately in young women and has a high incidence in Egypt. Successful completion of the studies proposed in this application will result in validated in vitro model systems that can be used for preclinical studies screening new therapeutics and may also identify biomarkers for IBC. Our intent is to improve the diagnosis and eventually the therapeutic management of this devastating cancer.