Targeting an MT1-MMP/MMP2 axis in melanoma by a novel MT1-MMP/MMP2 inhibitor

 描述（由申请人提供）：黑色素瘤仍然是最致命的皮肤癌形式，其五年预后仍然只有15%，强调缺乏有效的治疗。本发明的建议集中于通过新型MT 1-MMP/MMP特异性抑制剂靶向参与黑素瘤生长和进展的MT 1-MMP/MMP 2信号传导轴。MT 1-MMP是一种控制细胞周围蛋白水解的膜相关基质金属蛋白酶，是癌症中重要的、促进侵袭的、促肿瘤发生的MMP。我们最近发表的数据表明，MT 1-MMP表达的失调是一个早期事件，并在黑色素瘤进展过程中继续增加。MT 1-MMP表达与黑色素瘤患者预后不良相关，强调了MT 1-MMP在黑色素瘤发病机制中的关键作用。事实上，MT 1-MMP是黑色素瘤细胞转移所必需的，因为在原位小鼠黑色素瘤模型中剥夺MT 1-MMP的细胞不能形成远处转移。MT 1-MMP通过激活其靶点MMP 2影响细胞侵袭，而MMP 2也是MT 1-MMP维持RAC 1活性和促进MT 1-MMP依赖的细胞运动所必需的，突出了黑色素瘤中新的MT 1-MMP/MMP 2/RAC 1信号传导轴。然而，初步数据显示，虽然MT 1-MMP也控制细胞生长，但MMP 2不控制，这表明两种蛋白酶在黑色素瘤生长和传播中的不同作用。然而，我们的数据表明，这两种功能可以通过新型MT 1-MMP/MMP 2抑制剂ND-322在几种黑色素瘤细胞系中进行治疗靶向。ND-322减少黑素瘤细胞的迁移、侵袭和生长，类似于MT 1-MMP和/或MMP 2的特异性敲低。重要的是，虽然广谱MMP抑制剂缺乏特异性伴随着严重的副作用，例如由于靶向“好”和“坏”MMP而引起的肌肉骨骼疼痛和炎症，但ND-322在脑缺血的小鼠模型中显示出高耐受性。因此，我们假设特异性MT 1-MMP/MMP 2抑制剂ND-322代表了对患者现有治疗选择的安全、新的有效补充，其能够有效抑制黑色素瘤生长和转移。目的1将评估ND-322与维罗非尼（BRAF抑制剂）联合在BRAFV 600 E转基因小鼠黑素瘤模型中的功效，所述BRAFV 600 E转基因小鼠黑素瘤模型发展在遗传上和生物学上类似于人类疾病并且表达MT 1-MMP和MMP 2两者的病变。第二个目的是在原位小鼠黑色素瘤模型中，通过利用表达针对MT 1-MMP或MMP 2的特异性shRNA的人黑色素瘤细胞，表征ND-322下游的MT 1-MMP和MMP 2在调节黑色素瘤生长和转移中的作用。从这项工作中获得的知识将增强我们对黑色素瘤生长和进展机制的理解，同时为采用新型MMP抑制剂的治疗方法提供理论基础。

 DESCRIPTION (provided by applicant): Melanoma remains the deadliest form of skin cancer whose prognosis at five years is still only 15%, underscoring a lack of effective treatments. The present proposal focuses on the targeting of an MT1-MMP/MMP2 signaling axis involved in the growth and progression of melanoma by a novel MT1-MMP/MMP specific inhibitor. MT1-MMP is a membrane associated matrix metalloproteinase that controls pericellular proteolysis and is an important, invasion-promoting, pro-tumorigenic MMP in cancer. Our recently published data show that deregulation deregulation of MT1-MMP expression is an early event and continues to increase during melanoma progression. MT1-MMP expression is associated with poor melanoma patient outcome, underscoring a pivotal role of MT1-MMP in melanoma pathogenesis. Indeed, MT1-MMP is required for melanoma cells to metastasize, as cells deprived of MT1-MMP fail to form distant metastasis in an orthotopic mouse melanoma model. MT1-MMP affects cell invasion by activating its target MMP2, which isalso required by MT1-MMP to sustain RAC1 activity and promote MT1-MMP dependent cell motility, highlighting a novel MT1-MMP/MMP2/RAC1 signaling axis in melanoma. Yet, preliminary data show that while MT1-MMP also controls cell growth, MMP2 does not, pointing to distinct roles of the two proteases in melanoma growth and dissemination. Nevertheless, our data indicate both functions can be therapeutically targeted by the novel MT1-MMP/MMP2 inhibitor ND-322 in several melanoma cell lines. ND-322 diminishes melanoma cell migration, invasion and growth, similarly to specific knock down of MT1-MMP and/or MMP2. Importantly, while the lack of specificity of broad-spectrum MMP inhibitors have been accompanied by severe side effects, such as musculoskeletal pain and inflammation due to the targeting of both the "good" and the "bad" MMPs, ND-322 has shown high tolerability in mouse models of brain ischemia. Hence, we have hypothesized that the specific MT1-MMP/MMP2 inhibitor ND-322 represent a safe, novel valid addition to the current treatment options available to patients that is capable of effectivel inhibit melanoma growth and metastasis. Aim 1 will assess the efficacy of ND-322 in association with Vemurafenib (BRAF inhibitor) in a BRAFV600E transgenic mouse melanoma model that develops lesions that genetically and biologically resemble the human disease and that expresses both MT1-MMP and MMP2. The second aim will characterize, in an orthotopic mouse melanoma model the roles of MT1-MMP and MMP2 downstream of ND-322 in modulating melanoma growth and metastasis, by utilizing human melanoma cells expressing specific shRNAs against MT1-MMP or MMP2. The knowledge gained from this work will enhance our understanding of the mechanisms driving melanoma growth and progression while providing the rationale for a treatment approach employing a novel MMP inhibitor.