Department of Clinical Oncology, School of Medicine, Toho University, Tokyo, Japan.
Department of Obstetrics and Gynecology, School of Medicine, Ehime University, Ehime, Japan.
J Gene Med. 2019 Feb;21(2-3):e3064. doi: 10.1002/jgm.3064. Epub 2019 Feb 1.
A549 carrier cells infected with oncolytic adenovirus can induce complete tumor reduction of subcutaneous ovarian tumors but not intraperitoneal disseminated ovarian tumors. This appears to be a result of the insufficient antitumor effect of A549 carrier cells. Therefore, in the present study, we cloned a novel carrier cell with the aim of improving the antitumor effects.
Carrier cells infected with oncolytic adenovirus AdE3-midkine with a midkine promoter were cloned by limiting dilution. We examined the antitumor effects of these cells on subcutaneous and intraperitoneal OVHM ovarian tumors in a syngeneic mouse model. Biosafety tests were conducted in beagle dogs and rabbits.
We cloned EHMK-51-35 carrier cells with 10-fold higher antitumor effects compared to A549 carrier cells in vitro. EHMK-51-35 carrier cells co-infected with AdE3-midkine and Ad-mGM-CSF induced a 100% complete tumor reduction in subcutaneous tumors and a 60% reduction of intraperitoneal disseminated tumors. Single-dose acute toxicity test on beagle dogs with EHMK-51-35 carrier cells co-infected with AdE3-midkine and Ad-cGM-CSF showed no serious side effects. Biologically active adenoviruses were not detected in the blood, saliva, feces, urine or whole organs. In a chronic toxicity test, VX2 tumors in rabbits were injected five times with EHMK-51-35 carrier cells infected with AdE3-midkine and these rabbits showed no serious side effects.
Significant antitumor effects and safety of cloned EHMK-51-35 carrier cells were confirmed in intraperitoneal ovarian tumors and toxicity tests, respectively. These findings will be extended to preclinical efficacy studies using dogs and cats, with the aim of conducting human clinical trials on refractory solid tumors.
携带溶瘤腺病毒的 A549 载体细胞可诱导皮下卵巢肿瘤完全消退，但不能诱导腹腔内播散的卵巢肿瘤消退。这似乎是由于 A549 载体细胞的抗肿瘤作用不足所致。因此，本研究通过有限稀释克隆了一种新型载体细胞，旨在提高其抗肿瘤作用。
用 midkine 启动子克隆携带溶瘤腺病毒 AdE3-midkine 的载体细胞，在同基因小鼠模型中观察这些细胞对皮下和腹腔内 OVHM 卵巢肿瘤的抗肿瘤作用。在比格犬和兔中进行生物安全性测试。
我们克隆了 EHMK-51-35 载体细胞，与 A549 载体细胞相比，其在体外的抗肿瘤作用提高了 10 倍。EHMK-51-35 载体细胞共感染 AdE3-midkine 和 Ad-mGM-CSF 可使皮下肿瘤完全消退率达到 100%，腹腔内播散肿瘤消退率达到 60%。单次剂量急性毒性试验显示，EHMK-51-35 载体细胞共感染 AdE3-midkine 和 Ad-cGM-CSF 对比格犬无严重副作用。血液、唾液、粪便、尿液或全身器官均未检测到有生物活性的腺病毒。在慢性毒性试验中，将 EHMK-51-35 载体细胞感染 AdE3-midkine 后五次注入兔的 VX2 肿瘤，兔未出现严重副作用。
在腹腔内卵巢肿瘤中证实了克隆的 EHMK-51-35 载体细胞具有显著的抗肿瘤作用，在毒性试验中证实了其安全性。这些发现将扩展到使用犬和猫的临床前疗效研究，旨在为难治性实体瘤开展人体临床试验。
