Development of microRNA (miR)-based cell-targeted polymeric nanoparticles for myeloma therapy

项目摘要 多发性骨髓瘤（MM）是一种浆细胞在骨髓（BM）中定植的癌症， 使用新的有前途的治疗方式。这部分是由于（i）MM进展和耐药性 发育，（ii）通过BM微环境（BMME）保护MM细胞，和（iii）免疫逃避。因此，在本发明中， 迫切需要创新和更有效的治疗方法，特别是对于晚期疾病患者 对常规药剂无效。microRNA（miRs）在肿瘤的发生、发展和药物治疗中起着重要作用。 耐药性的各种人类癌症类型，包括MM，并提供了令人兴奋的机会，我们正在进行的 寻找新的和更有效的癌症治疗方法。我们最近记录到：（i）miR-30- 5 p家族 作为一种MM肿瘤抑制因子，靶向BCL 9，BCL 9是一种关键的Wnt/β-连环蛋白共激活因子， BM内皮细胞（BMEC），其促进MM细胞的BM定殖和增殖，（ii）miR-221/222 簇在来自对地塞米松无反应的患者的MM细胞中过表达， 作为MM癌基因，通过靶向CD 45 A和抑制细胞凋亡发挥作用，和（iii）miR-30 c-5 p和miR-221/222 在小鼠免疫细胞中表达，我们可以在移植的MM肿瘤中鉴定出小鼠巨噬细胞。 小鼠基于miR的治疗的主要挑战是需要安全有效的递送方法。除非 化学修饰或物理包封的miR在血液中不稳定，不易穿过细胞 膜的纳米颗粒（NPs）包括各种亚微米尺寸的大分子，其已经被用于制备纳米颗粒。 成功地作为包括miR在内的各种试剂的载体，使这些试剂能够到达细胞靶点 以前被认为是不可用的。兰格实验室已经成功地设计了一个多样化的聚合物库， NPs，其中一个范例7 C1 NP显示出无毒且有效地将siRNA递送至BMEC 对小鼠我的实验室随后表明，7 C1 NP制剂不仅可以将siRNA/miR递送到人， BMEC，但也对MM细胞以及鼠免疫细胞在体内。该项目的总体目标是 利用7 C1 NP递送系统（i）发现miR-30- 5 p的可能新靶点和作用 和miR-221/222在MM进展中的作用;以及（ii）探索这些聚合物包封的miR用于MM的潜力 通过miR-30- 5 p“替代疗法”靶向BMEC中的BCL 9并抑制BM中的MM生长的疗法，以及 (b)miR-221/222“反义（as）疗法”靶向MM细胞中的CD 40 A并增强细胞凋亡，同时废除 对来那度胺和硼替佐米的获得性耐药性，和（B）研究这些疗法对其他治疗的影响， 免疫细胞和MM相关的巨噬细胞极化。拟议的研究对公共卫生具有重要意义 因为它们将用来自患者的MM细胞系和MM细胞进行，并利用临床相关的 MM的小鼠异种移植物模型，其考虑了MM细胞与肿瘤细胞之间的异型相互作用。 BMME和他们的最终目标是改善患者的结果与更有效的治疗，减轻 不仅降低MM的总体治疗成本，而且降低潜在的其他血液恶性肿瘤的总体治疗成本。

Project Abstract Multiple myeloma (MM), a cancer of plasma cells that colonize the bone marrow (BM), remains incurable despite the use of new promising treatment modalities. This is partly due to (i) MM progression and drug resistance development, (ii) protection of MM cells by the BM microenvironment (BMME), and (iii) immune evasion. Thus, there is urgent need for innovative and more effective therapies, particularly for patients with advanced disease refractory to conventional agents. MicroRNAs (miRs) play critical roles in the initiation, progression, and drug resistance of various human cancer types, including MM, and are providing exciting opportunities in our ongoing search for novel and more effective cancer therapies. We recently documented that: (i) the miR-30-5p family serves as an MM-tumor suppressor targeting BCL9, a critical Wnt/-catenin co-activator, highly expressed in BM endothelial cells (BMECs), that promotes BM colonization and proliferation of MM cells, (ii) the miR-221/222 cluster is overexpressed in MM cells from patients who have become unresponsive to dexamethasone, and functions as an MM oncogene by targeting PUMA and inhibiting apoptosis, and (iii) miR-30c-5p and miR-221/222 are expressed in murine immune cells, and we can identify murine macrophages within MM tumors engrafted in mice. The main challenge for miR-based therapy is the need for safe and effective delivery methods. Unless chemically modified or physically encapsulated, miRs are unstable in the blood and do not easily cross the cell membrane. Nanoparticles (NPs) encompass a variety of submicron-sized macromolecules that have been used successfully as vehicles for various agents, including miRs, enabling these agents to reach cellular targets previously considered undruggable. The Langer lab has successfully engineered a diverse library of polymeric NPs, of which one exemplar, 7C1NP, was shown to be non-toxic and effective in delivering siRNAs to BMECs in mice. My lab subsequently showed that the 7C1NP formulation can deliver siRNAs/miRs not only to human BMECs but also to MM cells as well as murine immune cells in vivo. The overarching goal of this project is to take advantage of the 7C1NP delivery system to (i) uncover possible new targets of, and roles for, miR-30-5p and miR-221/222 in MM progression; and (ii) explore the potential of these polymer-encapsulated miRs for MM therapy via miR-30-5p “replacement therapy” to target BCL9 in BMECs, and inhibit MM growth in the BM, and (b) miR-221/222 “antisense (as) therapy” to target PUMA in MM cells and enhance apoptosis while abrogating acquired resistance to Lenalidomide, and Bortezomib, and (b) investigate the effect of these therapies on other immune cells and MM-associated macrophage polarization. The proposed studies are significant to public health in that they will be performed with MM cells lines and MM cells from patients and utilizing clinically relevant mouse xenograft models of MM that take in consideration the heterotypic interactions between MM cells and the BMME and their ultimate goal is to improve patient outcome with more efficacious therapies that alleviate suffering, and reduce the overall treatment cost of not only MM but potentially other hematologic malignancies.