Delgado-Bonet Pablo, Tomeo-Martín Beatriz Davinia, Ortiz-Díez Gustavo, Perisé-Barrios Ana Judith
Biomedical Research Unit, Universidad Alfonso X el Sabio, 28691 Villanueva de la Cañada, Spain.
Small Animal Surgery Service, Veterinary Teaching Hospital, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Vet Sci. 2022 Jun 9;9(6):285. doi: 10.3390/vetsci9060285.
Intravenous administration of oncolytic adenovirus (OAds) can be challenging, although various vehicles for the delivery of the virus to the tumor have been described. The efficacy of mesenchymal stem cells (MSCs) as a virus vehicle has been reported in mouse models and canine and human patients, but the actual action mechanism has never been described in patients. It is of importance to determine whether MSCs infected with OAds can reach the tumor and release the virus in a clinical setting. For this purpose, GFP-labeled MSCs were infected with an OAd and inoculated into a companion dog diagnosed with spontaneous lung carcinoma. Forty-eight hours later, the tumor was excised and analyzed microscopically by flow cytometry for GFP fluorescence detection, and a cellular culture was established. Peripheral blood samples were taken to quantify the oncolytic adenovirus by qRT-PCR. Green fluorescence cells detected in the cellular culture by microscopy and flow cytometry revealed 0.69% GFP-positive cells in the tumor. OAd in peripheral blood was confirmed by qRT-PCR during follow-up. For the first time, the tumoral-homing capacity of OAds infected-MSC has been confirmed in a clinical setting, helping to explain the clinical response mechanism, whose efficacy was previously reported in canine and human patients.
尽管已经描述了多种将病毒递送至肿瘤的载体,但静脉内给予溶瘤腺病毒(OAds)仍具有挑战性。间充质干细胞(MSCs)作为病毒载体的功效已在小鼠模型以及犬类和人类患者中得到报道,但尚未在患者中描述其实际作用机制。确定感染OAds的MSCs在临床环境中是否能够到达肿瘤并释放病毒至关重要。为此,用OAd感染绿色荧光蛋白(GFP)标记的MSCs,并将其接种到一只被诊断为自发性肺癌的伴侣犬体内。48小时后,切除肿瘤并通过流式细胞术进行显微镜分析以检测GFP荧光,同时建立细胞培养。采集外周血样本通过定量逆转录聚合酶链反应(qRT-PCR)对溶瘤腺病毒进行定量。通过显微镜和流式细胞术在细胞培养中检测到的绿色荧光细胞显示肿瘤中有0.69%的GFP阳性细胞。在随访期间通过qRT-PCR证实外周血中存在OAd。首次在临床环境中证实了感染OAds的MSCs的肿瘤归巢能力,这有助于解释临床反应机制,其疗效先前已在犬类和人类患者中报道。