Advanced Therapy Medicinal Product (ATMP) Department, Breast Cancer Research Center, Academic Center for Education, Culture and Research (ACECR), Motamed Cancer Institute, Tehran, Iran.
Advanced Therapy Medicinal Product (ATMP) Department, Breast Cancer Research Center, Academic Center for Education, Culture and Research (ACECR), Motamed Cancer Institute, Tehran, Iran; Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Int Immunopharmacol. 2024 Dec 25;143(Pt 2):113481. doi: 10.1016/j.intimp.2024.113481. Epub 2024 Oct 29.
Despite the recent advances in the diagnosis and treatment of breast cancer, triple-negative breast cancer (TNBC) remains a clinical challenge due to its aggressive nature and resistance to conventional therapies. Virotherapy has emerged as a promising cancer treatment strategy, leveraging the ability of viruses to specifically target and replicate in cancerous cells. This study evaluated the oncolytic potential of a combined therapeutic strategy, utilizing Newcastle disease virus (NDV) and Doxorubicin hydrochloride (Dox) both in vitro and in vivo.
The in vitro experiments involved exposing human and mouse TNBC cell lines (MDA-MB-231 and 4T1, respectively) to NDV and Dox, individually or in combination. Cell viability assays and flow cytometry analyses were conducted to assess the synergistic effects of NDV and Dox on regulating breast cancer cell behavior in vitro. Furthermore, the immune-stimulating potential of NDV was investigated by examining its effects on dendritic cell (DC) maturation using flow cytometry and T cell proliferation. The in vitro anti-tumor effects of NDV were examined in both parental and tamoxifen-resistant cancer cells to assess its efficacy against chemoresistance. Animal models of breast cancer were treated with NDV in combination with Dox. The body weight changes, tumor volume, and survival rates of the mice were monitored throughout the study. Histopathological analyses were conducted to evaluate the potential toxic effects of the treatments.
Based on the MTT results, NDV at optimal concentrations synergized the effect of Dox to reduce the viability of both MDA-MB231 and 4T1 cell lines (Isobologram combination index of less than 1). Additionally, individual treatment with NDV was able to significantly reduce the viability of patient-derived breast cancer cells, compared to the untreated control (P < 0.05) without affecting the cells of normal adjacent tissue. Furthermore, a combination of NDV and Dox significantly enhanced the percentage of early and late apoptotic cells in MDA-MB-231 (P < 0.0001) and late apoptotic cells in 4T1 (P < 0.0001), in comparison with individual treatment with these agents. Flow cytometry results showed that, compared to wild type MDA-MB-231 cells, NDV-infected MDA-MB-231 cells were better inducers of T cell proliferation and DC maturation as evidenced by increased proliferation index (P < 0.05) and elevated expression of CD1a, CD83, and CD86 (P < 0.0001), respectively. Moreover, co-treatment of both wild-type and (tamoxifen) TAM-resistant MCF-7/TAMR-1 cells with TAM and NDV significantly reduce the viability of the cancer cells (P < 0.0001). In tumor-bearing mice locally engrafted with 4T1 cells, combined treatment of NDV and Dox exhibited a marked reduction in median tumor volume compared to the control group, validating our in vitro findings on their synergistic anti-tumor effects. These findings suggest that combining NDV with Dox can effectively inhibit tumor progression and has the potential to reduce the dose, and consequently the toxic side-effects, of Dox in breast cancer therapy.
尽管乳腺癌的诊断和治疗最近取得了进展，但三阴性乳腺癌（TNBC）仍然是一个临床挑战，因为它具有侵袭性和对常规治疗的耐药性。溶瘤病毒治疗作为一种有前途的癌症治疗策略已经出现，利用病毒特异性靶向和复制癌细胞的能力。本研究评估了联合治疗策略的溶瘤潜力，该策略同时利用新城疫病毒（NDV）和盐酸多柔比星（Dox）进行体内和体外研究。
体外实验涉及将人源和鼠源 TNBC 细胞系（MDA-MB-231 和 4T1）暴露于 NDV 和 Dox 中，分别或联合使用。通过细胞活力测定和流式细胞术分析评估 NDV 和 Dox 对调节体外乳腺癌细胞行为的协同作用。此外，通过流式细胞术检测树突状细胞（DC）成熟和 T 细胞增殖，研究 NDV 的免疫刺激潜力。用 NDV 处理亲本和他莫昔芬耐药的乳腺癌细胞，评估其对化学耐药性的疗效，检验 NDV 在体外的抗肿瘤作用。用 NDV 联合 Dox 治疗乳腺癌动物模型。在整个研究过程中监测小鼠的体重变化、肿瘤体积和存活率。进行组织病理学分析以评估治疗的潜在毒性作用。
根据 MTT 结果，NDV 在最佳浓度下与 Dox 协同作用，降低 MDA-MB231 和 4T1 细胞系的活力（低于 1 的等药效线组合指数）。此外，与未处理的对照组相比，NDV 单独处理能够显著降低患者来源的乳腺癌细胞的活力（P < 0.05），而对正常相邻组织的细胞没有影响。此外，与单独使用这些药物相比，NDV 和 Dox 的联合使用显著增加了 MDA-MB-231 中的早期和晚期凋亡细胞的百分比（P < 0.0001）和 4T1 中的晚期凋亡细胞的百分比（P < 0.0001）。流式细胞术结果表明，与野生型 MDA-MB-231 细胞相比，感染 NDV 的 MDA-MB-231 细胞能更好地诱导 T 细胞增殖和 DC 成熟，表现为增殖指数增加（P < 0.05）和 CD1a、CD83 和 CD86 的表达升高（P < 0.0001）。此外，用 TAM 和 NDV 联合处理野生型和（他莫昔芬）TAM 耐药 MCF-7/TAMR-1 细胞，可显著降低癌细胞的活力（P < 0.0001）。在局部植入 4T1 细胞的荷瘤小鼠中，与对照组相比，NDV 和 Dox 的联合治疗显著降低了肿瘤的中位体积，验证了我们在体外研究中关于其协同抗肿瘤作用的发现。这些发现表明，联合使用 NDV 和 Dox 可以有效抑制肿瘤进展，并有可能降低乳腺癌治疗中 Dox 的剂量，从而降低其毒性副作用。
