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

项目摘要 多发性骨髓瘤(MM)是一种定植于骨髓(BM)的浆细胞癌，尽管如此，仍无法治愈 使用新的有希望的治疗方式。这在一定程度上是由于(I)MM的进展和耐药性 (Ii)骨髓微环境(BMME)对MM细胞的保护，以及(Iii)免疫逃避。因此， 迫切需要创新和更有效的治疗方法，特别是对晚期疾病患者。 对传统药剂不起作用。MicroRNAs(MiRs)在肿瘤的发生、发展和药物治疗中起着重要作用。 抵抗各种人类癌症类型，包括多发性骨髓瘤，并在我们正在进行的 寻找新的、更有效的癌症疗法。我们最近记录了：(I)miR-30-5P系列 作为针对BCL9的MM-肿瘤抑制因子，BCL9是一种关键的WNT/-连环蛋白共激活因子，在 促进骨髓细胞定植和增殖的骨髓内皮细胞(BMECs)，(Ii)miR-221/222 在对地塞米松无效的患者的MM细胞中，CLUSTER过表达，并且 通过靶向PUMA和抑制细胞凋亡作为MM癌基因，以及(Iii)miR-30c-5p和miR-221/222 在小鼠免疫细胞中表达，我们可以在移植的MM肿瘤中识别小鼠巨噬细胞 老鼠。基于miR的治疗的主要挑战是需要安全有效的给药方法。除非 经过化学修饰或物理包裹的MIR在血液中不稳定，不容易穿过细胞 薄膜。纳米粒子(NPs)包括各种已被使用的亚微米大分子 成功地作为各种药物的载体，包括MIR，使这些药物能够达到细胞靶标 以前被认为是无法下药的。兰格实验室已经成功地设计出一种不同的聚合物文库 NPS，其中一个样本7C1NP被证明是无毒的，并且在向BMEC运送siRNAs方面有效 在老鼠身上。我的实验室随后发现，7C1NP配方不仅可以将siRNAs/miRs传递给人类 BMECs也能在体内对MM细胞以及小鼠免疫细胞起作用。这个项目的首要目标是 利用7C1NP递送系统：(I)发现miR-30-5P可能的新目标和作用 和miR-221/222在多发性骨髓瘤进展中的作用；以及(Ii)探索这些聚合物包裹的miR在多发性骨髓瘤治疗中的潜力 通过miR-30-5P“替代疗法”靶向BMECs中的BCL9，并抑制骨髓中MM的生长，以及 (B)以多发性骨髓瘤细胞中的PUMA为靶点的miR-221/222“反义(AS)疗法”，在去除PUMA的同时促进细胞凋亡 对Lenalidomide和Bortezomib的获得性耐药性，以及(B)调查这些疗法对其他 免疫细胞和MM相关巨噬细胞极化。建议的研究对公众健康具有重要意义。 这是因为它们将用患者的MM细胞系和MM细胞进行，并利用临床上相关的 小鼠异种移植多发性骨髓瘤模型，考虑了多发性骨髓瘤细胞和肿瘤细胞之间的异型相互作用。 BMME和他们的最终目标是通过更有效的治疗来改善患者的预后 不仅降低了多发性骨髓瘤的总体治疗成本，还降低了潜在的其他恶性血液疾病的治疗成本。

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.