Local and systemic control of multiple myeloma colonization and growth by MMP-13

总结 尽管医学进步，多发性骨髓瘤仍然是致命的疾病。这些机制如何支撑 骨髓瘤在局部骨微环境中进展，骨髓瘤通过该微环境定植于骨骼， 产生多个疼痛的溶骨性病变需要紧急解决。基质金属蛋白酶 是通过调节细胞因子和生长因子来调节肿瘤-骨相互作用的关键调节因子 生物利用度/活性。我们小组的新数据已经确定，在这种疾病的人类样本中， MMP-13由骨髓瘤细胞和骨微环境细胞高度表达， 造骨细胞体内分析表明，当MMP-13从宿主中基因消除时， 在一个室中，总生存率显著增加，同时骨髓瘤诱导的 骨病我们还发现骨髓瘤来源的外泌体可以促进骨骼定植， 富含MMP-13。此外，一种新型高选择性MMP-13抑制剂的初步数据 在体外和体内显著限制骨髓瘤生长，强调了MMP-13活性在驱动骨髓瘤生长中的作用。 多发性骨髓瘤基于这些初步发现，我们假设MMP-13有助于多种 骨髓瘤进展。我们将测试我们的假设：1）定义肿瘤和成骨细胞衍生的作用， MMP-13在多发性骨髓瘤进展中的作用我们将使用CRISPR/cDNA过表达方法， 在使用特异性Cre重组酶驱动的启动子的同时操纵骨髓瘤细胞系中MMP-13的水平 消除成骨细胞MMP-13的表达。然后，我们将测试MMP-13的存在与否， 在一个或两个隔室中，骨髓瘤生长和相关的骨疾病在两个单独的隔室中起作用。 体内模型（5 TGM 1和U266）。我们还将探索MMP-13的作用机制与前期工作 表明转化生长因子β（TGFβ）活性的调节。2)确定骨髓瘤的作用 衍生的外泌体，特别是外泌体MMP-13在疾病的骨骼定殖中的作用。我们还将 检查外泌体MMP-13是否可以识别处于高风险的郁积型多发性骨髓瘤患者（n=200）， 使用蛋白质组学技术进展为活动性疾病。3)确定选择性MMP-13的功效 使用CD 138分离的骨髓瘤细胞限制多发性骨髓瘤活力的抑制剂， 在新的离体高通量平台和临床相关的体内模型中， 疾病基于预期的结果，探讨MMP-13在多发性骨髓瘤进展中的作用将有助于我们更好地了解MMP-13在多发性骨髓瘤进展中的作用。 揭示了许多新的见解，从而为选择性MMP-13的翻译提供了强有力的理论基础 我们预测，由于骨骼肌的限制， MMP-13的表达。

SUMMARY Despite medical advances, multiple myeloma remains a fatal disease. The mechanisms underpinning how myeloma progresses in the local bone microenvironment and through which myeloma colonizes the skeleton to generate multiple painful osteolytic lesions needs to be addressed urgently. Matrix metalloproteinases (MMPs) are key regulators of tumor-bone interaction via the regulation of cytokine and growth factor bioavailability/activity. Emerging data from our group has identified that in human specimens of the disease, MMP-13 is highly expressed by both the myeloma cells and the cells of the bone microenvironment namely, bone building osteoblasts. In vivo analyses show that when MMP-13 is genetically ablated from the host compartment there is a significant increase in overall survival and a concomitant decrease in myeloma induced bone disease. We also have identified that myeloma derived exosomes can promote the skeletal colonization of myeloma and, are rich in MMP-13. Further, preliminary data with a novel highly selective MMP-13 inhibitor significantly limits myeloma growth in vitro and in vivo underscoring the role for MMP-13 activity in driving multiple myeloma. Based on these preliminary findings we hypothesize that MMP-13 contributes to multiple myeloma progression. We will test our hypothesis by; 1) Defining the role of tumor and osteoblast derived MMP-13 in multiple myeloma progression. We will use CRISPR/cDNA overexpression approaches to manipulate the levels of MMP-13 in myeloma cell lines while using specific Cre-recombinase driven promoters to eliminate MMP-13 expression by osteoblasts. We will then test whether the presence or absence of MMP-13 in one or both compartments contributes to myeloma growth and associated bone disease in two separate in vivo models (5TGM1 and U266). We will also explore MMP-13 mechanisms of action with preliminary work pointing to regulation of transforming growth factorβ (TGFβ) activity. 2) Determining the role of myeloma derived exosomes and specifically exosomal MMP-13 in the skeletal colonization of the disease. We will also examine whether exosomal MMP-13, can identify smoldering multiple myeloma patients (n=200) at high-risk of progression to active disease using proteomic techniques. 3) Identifying the efficacy of a selective MMP-13 inhibitor in limiting multiple myeloma viability using CD138 isolated myeloma cells obtained from newly diagnosed patients in a novel ex vivo high throughput platform and clinically relevant in vivo models of the disease. Based on the anticipated results, interrogating the role MMP-13 in multiple myeloma progression will reveal a number of novel insights thereby providing a strong rationale for the translation of selective MMP-13 inhibitors to the clinic that we predict would have limited off-target effects due to the restricted skeletal expression of MMP-13.