VandenHeuvel Sabrina N, Chau Eric, Mohapatra Arpita, Dabbiru Sameera, Roy Sanjana, O'Connell Cailin, Kamat Aparna, Godin Biana, Raghavan Shreya A
Department of Biomedical Engineering, Texas A&M University, 3120 TAMU, College Station, Texas 77843, United States.
Department of Nanomedicine, Houston Methodist Research Institute, 6670 Bertner Avenue, Houston, Texas 77030, United States.
ACS Appl Bio Mater. 2024 Dec 16;7(12):7871-7882. doi: 10.1021/acsabm.4c00076. Epub 2024 Apr 1.
Most ovarian carcinoma (OvCa) patients present with advanced disease at the time of diagnosis. Malignant, metastatic OvCa is invasive and has poor prognosis, exposing the need for improved therapeutic targeting. High CD47 (OvCa) and SIRPα (macrophage) expression has been linked to decreased survival, making this interaction a significant target for therapeutic discovery. Even so, previous attempts have fallen short, limited by CD47 antibody specificity and efficacy. Macrophages are an important component of the OvCa tumor microenvironment and are manipulated to aid in cancer progression via CD47-SIRPα signaling. Thus, we have leveraged lipid-based nanoparticles (LNPs) to design a therapy uniquely situated to home to phagocytic macrophages expressing the SIRPα protein in metastatic OvCa. CD47-SIRPα presence was evaluated in patient histological sections using immunohistochemistry. 3D tumor spheroids generated on a hanging drop array with OVCAR3 high-grade serous OvCa and THP-1-derived macrophages created a representative model of cellular interactions involved in metastatic OvCa. Microfluidic techniques were employed to generate LNPs encapsulating SIRPα siRNA (siSIRPα) to affect the CD47-SIRPα signaling between the OvCa and macrophages. siSIRPα LNPs were characterized for optimal size, charge, and encapsulation efficiency. Uptake of the siSIRPα LNPs by macrophages was assessed by Incucyte. Following 48 h of 25 nM siSIRPα treatment, OvCa/macrophage heterospheroids were evaluated for SIRPα knockdown, platinum chemoresistance, and invasiveness. OvCa patient tumors and heterospheroids expressed CD47 and SIRPα. Macrophages in OvCa spheroids increased carboplatin resistance and invasion, indicating a more malignant phenotype. We observed successful LNP uptake by macrophages causing significant reduction in gene and protein expressions and subsequent reversal of pro-tumoral alternative activation. Disrupting CD47-SIRPα interactions resulted in sensitizing OvCa/macrophage heterospheroids to platinum chemotherapy and reversal of cellular invasion outside of heterospheroids. Ultimately, our results strongly indicate the potential of using LNP-based nanoimmunotherapy to reduce malignant progression of ovarian cancer.
大多数卵巢癌（OvCa）患者在诊断时已处于疾病晚期。恶性转移性OvCa具有侵袭性且预后较差，这凸显了改进治疗靶点的必要性。高CD47（OvCa）和信号调节蛋白α（SIRPα，巨噬细胞）表达与生存率降低有关，使得这种相互作用成为治疗探索的重要靶点。即便如此，先前的尝试均未成功，受限于CD47抗体的特异性和疗效。巨噬细胞是OvCa肿瘤微环境的重要组成部分，并通过CD47-SIRPα信号传导被操控以促进癌症进展。因此，我们利用脂质纳米颗粒（LNP）设计了一种独特的疗法，该疗法能够靶向转移性OvCa中表达SIRPα蛋白的吞噬性巨噬细胞。使用免疫组织化学方法在患者组织切片中评估CD47-SIRPα的存在情况。在悬滴阵列上利用高级别浆液性OvCa的OVCAR3细胞和THP-1来源的巨噬细胞生成的三维肿瘤球体，构建了一个转移性OvCa中细胞相互作用的代表性模型。采用微流控技术生成包裹SIRPα小干扰RNA（siSIRPα）的LNP，以影响OvCa和巨噬细胞之间的CD47-SIRPα信号传导。对siSIRPα LNP的最佳尺寸、电荷和包封效率进行了表征。通过Incucyte评估巨噬细胞对siSIRPα LNP的摄取情况。在25 nM siSIRPα处理48小时后，评估OvCa/巨噬细胞异质球体的SIRPα敲低、铂类化疗耐药性和侵袭性。OvCa患者肿瘤和异质球体表达CD47和SIRPα。OvCa球体中的巨噬细胞增加了卡铂耐药性和侵袭性，表明其具有更恶性的表型。我们观察到巨噬细胞成功摄取LNP，导致基因和蛋白表达显著降低，并随后逆转了促肿瘤的替代激活。破坏CD47-SIRPα相互作用导致OvCa/巨噬细胞异质球体对铂类化疗敏感，并逆转了异质球体外部的细胞侵袭。最终，我们的结果有力地表明了基于LNP的纳米免疫疗法降低卵巢癌恶性进展的潜力。
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