Adenovirus-mediated efficient gene transfer into cultured three-dimensional organoids.
Three-dimensional organoids have been recently established from various tissue-specific progenitors (such as intestinal stem cells), induced pluripotent stem cells, or embryonic stem cells. These cultured self-sustaining stem cell-based organoids may become valuable systems to study the roles of tissue-specific stem cells in tissue genesis and disease development. It is thus conceivable that effective genetic manipulations in such organoids may allow us to reconstruct disease processes and/or develop novel therapeutics. Recombinant adenoviruses are one of the most commonly used viral vectors for in vitro and in vivo gene deliveries. In this study, we investigate if adenoviruses can be used to effectively deliver transgenes into the cultured "mini-gut" organoids derived from intestinal stem cells. Using adenoviral vectors that express fluorescent proteins, we demonstrate that adenoviruses can effectively deliver transgenes into the cultured 3-D "mini-gut" organoids. The transgene expression can last at least 10 days in the cultured organoids. As a proof-of-principle experiment, we demonstrate that adenovirus-mediated noggin expression effectively support the survival and self-renewal of mini-gut organoids, while adenovirus-mediated expression of BMP4 inhibits the self-sustainability and proliferation of the organoids. Thus, our results strongly suggest that adenovirus vectors can be explored as effective gene delivery vehicles to introduce genetic manipulations in 3-D organoids.
Department of Oncology and Southwest Cancer Center, the Affiliated Southwest Hospital, Third Military Medical University, Chongqing, China; Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America.
Ning
N
Wang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Hongyu
H
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Bing-Qiang
BQ
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Wei
W
Liu
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Department of Surgery, Affiliated Zhongnan Hospital of Wuhan University, Wuhan, China.
Zhonglin
Z
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Min
M
Qiao
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Hongmei
H
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America.
Fang
F
Deng
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Ningning
N
Wu
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Xian
X
Chen
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Sheng
S
Wen
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Junhui
J
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Department of Orthopaedic Surgery, Xiang-Ya Hospital of Central South University, Changsha, China.
Zhan
Z
Liao
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Qian
Q
Zhang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Zhengjian
Z
Yan
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Liangjun
L
Yin
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; School of Bioengineering, Chongqing University, Chongqing, China.
Jixing
J
Ye
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Youlin
Y
Deng
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America.
Hue H
HH
Luu
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America.
Rex C
RC
Haydon
Department of Oncology and Southwest Cancer Center, the Affiliated Southwest Hospital, Third Military Medical University, Chongqing, China.
Houjie
H
Liang
Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of America; Ministry of Education Key Laboratory of Clinical Diagnostic Medicine, and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
Tong-Chuan
TC
He
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017077:Culture Media, Conditioned; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D007422:Intestines / Q000166:cytology*; D051379:Mice
D017077:Culture Media, Conditioned