Adenovirus-mediated stem cell leukemia gene transfer induces rescue of interstitial cells of Cajal in ICC-loss mice.
OBJECTIVE
Interaction of c-Kit and its ligand stem cell factor (SCF) is necessary for appropriate development and survival of interstitial cells of Cajal (ICC) in the intestine. Blockade of c-Kit will cause ICC loss in vivo. Stem cell leukemia (SCL) gene acts as a positive regulator of upstream transcription of c-Kit expression. This study aimed to explore whether the restoration of c-Kit expression promoted by SCL gene transfer could rescue ICC in vivo.


METHODS
A modified ICC-loss mouse model was created by continual administration of anti-c-Kit antibody (ACK2) to obtain a steady status of ICC loss, and a recombinant adenovirus vector containing SCL gene (Ad-SCL) was designed to rescue ICC in these mice. Western blot analysis and immunofluorescence labeling assays were performed to analyze the SCL and c-Kit expression in vitro and in vivo. The distribution and configuration of ICC were observed with immunohistochemistry and electromicroscope.


RESULTS
Western blot analysis and immunofluorescence labeling assays showed that SCL gene was successfully delivered to cultured HeLa and ICC cells in vitro. Moreover, significantly increased c-Kit expression could be detected in the colon of Ad-SCL-infected ICC-loss mice. Furthermore, rescue of the ICC network and ICC with typical ultrastructural features could be detected in Ad-SCL-infected ICC-loss mice at day 37.


CONCLUSIONS
Ad-SCL was able to enhance c-Kit expression, reactivate the c-Kit/SCF pathway, and rescue ICC in ICC-loss mice. Since loss and defects of ICC are associated with many human gut motility disorders, Ad-SCL may be of potential use in gene therapy of these patients.
Li
Fan
F
Department of General Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China.
Zhang
Lin
L
Li
Chunxue
C
Ni
Bing
B
Wu
Yuzhang
Y
Huang
Ying
Y
Zhang
Guangjun
G
Wang
Li
L
Zhang
Anping
A
He
Yujun
Y
Fu
Tao
T
Tong
Weidong
W
Liu
Baohua
B
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D048430:Cell Shape; D003106:Colon / Q000473:pathology / Q000648:ultrastructure; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D056885:Interstitial Cells of Cajal / Q000473:pathology* / Q000648:ultrastructure; D007938:Leukemia / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D013194:Staining and Labeling; D013234:Stem Cells / Q000378:metabolism*
D051792:Basic Helix-Loop-Helix Proteins; D019009:Proto-Oncogene Proteins c-kit