TY - JOUR
T1 - Functional analysis and drug response to zinc and D-penicillamine in stable ATP7B mutant hepatic cell lines
AU - Chandhok, Gursimran
AU - Horvath, Judit
AU - Aggarwal, Annu
AU - Bhatt, Mohit
AU - Zibert, Andree
AU - Schmidt, Hartmut H.J.
PY - 2016/4/28
Y1 - 2016/4/28
N2 - AIM: To study the effect of anti-copper treatment for survival of hepatic cells expressing different ATP7B mutations in cell culture. METHODS: The most common Wilson disease (WD) mutations p.H1069Q, p.R778L and p.C271∗, found in the ATP7B gene encoding a liver copper transporter, were studied. The mutations represent major genotypes of the United States and Europe, China, and India, respectively. A human hepatoma cell line previously established to carry a knockout of ATP7B was used to stably express WD mutants. mRNA and protein expression of mutant ATP7B , survival of cells, apoptosis, and protein trafficking were determined. RESULTS: Low temperature increased ATP7B protein expression in several mutants. Intracellular ATP7B localization was significantly impaired in the mutants. Mutants were classified as high, moderate, and no survival based on their viability on exposure to toxic copper. Survival of mutant p.H1069Q and to a lesser extent p.C271∗ improved by D-penicillamine (DPA) treatment, while mutant p.R778L showed a pronounced response to zinc (Zn) treatment. Overall, DPA treatment resulted in higher cell survival as compared to Zn treatment; however, only combined Zn + DPA treatment fully restored cell viability. CONCLUSION: The data indicate that the basic impact of a genotype might be characterized by analysis of mutant hepatic cell lines.
AB - AIM: To study the effect of anti-copper treatment for survival of hepatic cells expressing different ATP7B mutations in cell culture. METHODS: The most common Wilson disease (WD) mutations p.H1069Q, p.R778L and p.C271∗, found in the ATP7B gene encoding a liver copper transporter, were studied. The mutations represent major genotypes of the United States and Europe, China, and India, respectively. A human hepatoma cell line previously established to carry a knockout of ATP7B was used to stably express WD mutants. mRNA and protein expression of mutant ATP7B , survival of cells, apoptosis, and protein trafficking were determined. RESULTS: Low temperature increased ATP7B protein expression in several mutants. Intracellular ATP7B localization was significantly impaired in the mutants. Mutants were classified as high, moderate, and no survival based on their viability on exposure to toxic copper. Survival of mutant p.H1069Q and to a lesser extent p.C271∗ improved by D-penicillamine (DPA) treatment, while mutant p.R778L showed a pronounced response to zinc (Zn) treatment. Overall, DPA treatment resulted in higher cell survival as compared to Zn treatment; however, only combined Zn + DPA treatment fully restored cell viability. CONCLUSION: The data indicate that the basic impact of a genotype might be characterized by analysis of mutant hepatic cell lines.
KW - Atp7b
KW - D-penicillamine
KW - Mutations
KW - Therapy
KW - Wilson disease
KW - Zinc
UR - http://www.scopus.com/inward/record.url?scp=84969930720&partnerID=8YFLogxK
U2 - 10.3748/wjg.v22.i16.4109
DO - 10.3748/wjg.v22.i16.4109
M3 - Article
C2 - 27122662
AN - SCOPUS:84969930720
SN - 1007-9327
VL - 22
SP - 4109
EP - 4119
JO - World Journal of Gastroenterology
JF - World Journal of Gastroenterology
IS - 16
ER -
