br To prepare CAR NK cells
To prepare CAR-NK Relebactam for the clinical study, blood samples were collected from a patient (100 mL) or haploidentical family do-nors (200 mL) to isolate PBMCs. NK cells were expanded from PBMCs as described above in G-Rex 100 vessels (Wilson Wolf Manufacturing), supplemented with 1% human AB serum (Gemini Bio-Products, Calabasas, CA, USA) and 50 IU/mL human recombi-nant IL-2 (Beijing Four Rings Biopharmaceutical, China). NKG2Dp CAR NK cells were prepared as described above and in the Supple-mental Information (Figure S1) and administered i.p. via a perito-neal access catheter to patients. Percutaneous injection of CAR-NK cells was performed under real-time ultrasound (SIEMENS Acuson S2000) guidance with a 3 to 6 MHz probe. The patient was placed in the left lateral position, and the ultrasound scan was performed on the right anterior iliac crest. Two tumor sties without major blood vessels were selected for the injection, 5 108 to 2 109 CAR NK cells in 50–70 mL saline each time, through the seventh and eighth rib intercostal space with a 19G disposable puncture needle. To evaluate the antitumor effects of NKG2D CAR-NK cell therapy on malignant ascites, peritoneal lavage fluid was collected through a catheter. Tumor cells in ascites were analyzed by flow cytometry for the presence of EpCAM-posi-tive cells.
Data are presented as mean ± standard deviation (SD). All statistics were performed with GraphPad Prism 7. p values < 0.05 were consid-ered significant.
Supplemental Information can be found online at https://doi.org/10.
CONFLICTS OF INTEREST
The costs of the study were partially covered by Youshan Biomedical Co., Ltd. (Hangzhou, China). The study sponsors had no involvement in study design, collection and interpretation of data, writing the report, and the decision to submit the report for publication. All other authors in the Third Affiliated Hospital of Guangzhou Medical Uni-versity and Guangzhou Regenerative Medicine, Health-Guangdong Laboratory (GRMH-GDL), Guangzhou, and the Affiliated Hangzhou First’s People Hospital, Zhejiang University School of Medicine, Hangzhou, China, have declared that there are no financial conflicts of interest related to this work.
This work was supported by the Science and Technology Planning Projects of Guangdong Province, China (grants 2016B090918130 and 2016ZC0142) and the General Project of Science and Technology from Guangzhou City (grant 201707010234).
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Molecular and Cellular Endocrinology
journal homepage: www.elsevier.com/locate/mce
Adrenal androgens rescue prostatic dihydrotestosterone production and growth of prostate cancer cells after castration
Yue Wua,∗, Li Tangb, Gissou Azabdaftaric, Elena Popa, Gary J. Smitha a Department of Urology, Roswell Park Comprehensive Cancer Center, Buﬀalo, NY 14263, USA
b Department of Cancer Prevention and Control, Roswell Park Comprehensive Cancer Center, Buﬀalo, NY 14263, USA
c Department of Pathology, Roswell Park Comprehensive Cancer Center, Buﬀalo, NY 14263, USA
Intracrine androgen metabolism
Adrenal androgens dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEAS) are potential substrates for intracrine production of testosterone (T) and dihydrotestosterone (DHT), or directly to DHT, by prostate cancer (PCa) cells. Production of DHT from DHEAS and DHEA, and the role of steroid sulfatase (STS), were evaluated ex vivo using fresh human prostate tissue and in vitro using human PCa cell lines. STS was expressed in benign prostate tissue and PCa tissue. DHEAS at a physiological concentration was converted to DHT in prostate tissue and PCa cell lines, which was STS-dependent. DHEAS activation of androgen receptor (AR) and stimulation of PCa cell growth were STS-dependent. DHEA at a physiological concentration was not converted to DHT ex vivo and in vitro, but stimulated in vivo tumor growth of the human PCa cell line, VCaP, in castrated mice. The findings suggest that targeting metabolism of DHEAS and DHEA may enhance androgen deprivation therapy.