Mechanisms of CFTR Functional Variants That Impair Regulated Bicarbonate Permeation and Increase Risk for Pancreatitis but Not for Cystic Fibrosis

Jessica LaRusch, Jinsei Jung, Ignacio J. General, Michele D. Lewis, Hyun Woo Park, Randall E. Brand, Andres Gelrud, Michelle A. Anderson, Peter A. Banks, Darwin Conwell, Christopher Lawrence, Joseph Romagnuolo, John Baillie, Samer Alkaade, Gregory Cote, Timothy B. Gardner, Stephen T. Amann, Adam Slivka, Bimaljit Sandhu, Amy Aloe & 6 others Michelle L. Kienholz, Dhiraj Yadav, M. Michael Barmada, Ivet Bahar, Min Goo Lee, David C. Whitcomb

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev) cause complete loss of CFTR function and result in cystic fibrosis (CF), a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize that those CFTR mutations that disrupt the WNK1-SPAK activation mechanisms cause a selective, bicarbonate defect in channel function (CFTRBD) affecting organs that utilize CFTR for bicarbonate secretion (e.g. the pancreas, nasal sinus, vas deferens) but do not cause typical CF. To understand the structural and functional requirements of the CFTR bicarbonate-preferring channel, we (a) screened 984 well-phenotyped pancreatitis cases for candidate CFTRBD mutations from among 81 previously described CFTR variants; (b) conducted electrophysiology studies on clones of variants found in pancreatitis but not CF; (c) computationally constructed a new, complete structural model of CFTR for molecular dynamics simulation of wild-type and mutant variants; and (d) tested the newly defined CFTRBD variants for disease in non-pancreas organs utilizing CFTR for bicarbonate secretion. Nine variants (CFTR R74Q, R75Q, R117H, R170H, L967S, L997F, D1152H, S1235R, and D1270N) not associated with typical CF were associated with pancreatitis (OR 1.5, p = 0.002). Clones expressed in HEK 293T cells had normal chloride but not bicarbonate permeability and conductance with WNK1-SPAK activation. Molecular dynamics simulations suggest physical restriction of the CFTR channel and altered dynamic channel regulation. Comparing pancreatitis patients and controls, CFTRBD increased risk for rhinosinusitis (OR 2.3, p<0.005) and male infertility (OR 395, p≪0.0001). WNK1-SPAK pathway-activated increases in CFTR bicarbonate permeability are altered by CFTRBD variants through multiple mechanisms. CFTRBD variants are associated with clinically significant disorders of the pancreas, sinuses, and male reproductive system.

Original languageEnglish
Article numbere1004376
JournalPLoS Genetics
Volume10
Issue number7
DOIs
Publication statusPublished - 2014 Jan 1

Fingerprint

pancreatitis
cystic fibrosis
Bicarbonates
bicarbonates
bicarbonate
Cystic Fibrosis
Pancreatitis
paranasal sinuses
Pancreas
mutation
Permeability
permeability
male reproductive system
molecular dynamics
Paranasal Sinuses
Molecular Dynamics Simulation
pancreas
secretion
Mutation
Chlorides

All Science Journal Classification (ASJC) codes

  • Ecology, Evolution, Behavior and Systematics
  • Molecular Biology
  • Genetics
  • Genetics(clinical)
  • Cancer Research

Cite this

LaRusch, Jessica ; Jung, Jinsei ; General, Ignacio J. ; Lewis, Michele D. ; Park, Hyun Woo ; Brand, Randall E. ; Gelrud, Andres ; Anderson, Michelle A. ; Banks, Peter A. ; Conwell, Darwin ; Lawrence, Christopher ; Romagnuolo, Joseph ; Baillie, John ; Alkaade, Samer ; Cote, Gregory ; Gardner, Timothy B. ; Amann, Stephen T. ; Slivka, Adam ; Sandhu, Bimaljit ; Aloe, Amy ; Kienholz, Michelle L. ; Yadav, Dhiraj ; Barmada, M. Michael ; Bahar, Ivet ; Lee, Min Goo ; Whitcomb, David C. / Mechanisms of CFTR Functional Variants That Impair Regulated Bicarbonate Permeation and Increase Risk for Pancreatitis but Not for Cystic Fibrosis. In: PLoS Genetics. 2014 ; Vol. 10, No. 7.
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abstract = "CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev) cause complete loss of CFTR function and result in cystic fibrosis (CF), a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize that those CFTR mutations that disrupt the WNK1-SPAK activation mechanisms cause a selective, bicarbonate defect in channel function (CFTRBD) affecting organs that utilize CFTR for bicarbonate secretion (e.g. the pancreas, nasal sinus, vas deferens) but do not cause typical CF. To understand the structural and functional requirements of the CFTR bicarbonate-preferring channel, we (a) screened 984 well-phenotyped pancreatitis cases for candidate CFTRBD mutations from among 81 previously described CFTR variants; (b) conducted electrophysiology studies on clones of variants found in pancreatitis but not CF; (c) computationally constructed a new, complete structural model of CFTR for molecular dynamics simulation of wild-type and mutant variants; and (d) tested the newly defined CFTRBD variants for disease in non-pancreas organs utilizing CFTR for bicarbonate secretion. Nine variants (CFTR R74Q, R75Q, R117H, R170H, L967S, L997F, D1152H, S1235R, and D1270N) not associated with typical CF were associated with pancreatitis (OR 1.5, p = 0.002). Clones expressed in HEK 293T cells had normal chloride but not bicarbonate permeability and conductance with WNK1-SPAK activation. Molecular dynamics simulations suggest physical restriction of the CFTR channel and altered dynamic channel regulation. Comparing pancreatitis patients and controls, CFTRBD increased risk for rhinosinusitis (OR 2.3, p<0.005) and male infertility (OR 395, p≪0.0001). WNK1-SPAK pathway-activated increases in CFTR bicarbonate permeability are altered by CFTRBD variants through multiple mechanisms. CFTRBD variants are associated with clinically significant disorders of the pancreas, sinuses, and male reproductive system.",
author = "Jessica LaRusch and Jinsei Jung and General, {Ignacio J.} and Lewis, {Michele D.} and Park, {Hyun Woo} and Brand, {Randall E.} and Andres Gelrud and Anderson, {Michelle A.} and Banks, {Peter A.} and Darwin Conwell and Christopher Lawrence and Joseph Romagnuolo and John Baillie and Samer Alkaade and Gregory Cote and Gardner, {Timothy B.} and Amann, {Stephen T.} and Adam Slivka and Bimaljit Sandhu and Amy Aloe and Kienholz, {Michelle L.} and Dhiraj Yadav and Barmada, {M. Michael} and Ivet Bahar and Lee, {Min Goo} and Whitcomb, {David C.}",
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month = "1",
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doi = "10.1371/journal.pgen.1004376",
language = "English",
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LaRusch, J, Jung, J, General, IJ, Lewis, MD, Park, HW, Brand, RE, Gelrud, A, Anderson, MA, Banks, PA, Conwell, D, Lawrence, C, Romagnuolo, J, Baillie, J, Alkaade, S, Cote, G, Gardner, TB, Amann, ST, Slivka, A, Sandhu, B, Aloe, A, Kienholz, ML, Yadav, D, Barmada, MM, Bahar, I, Lee, MG & Whitcomb, DC 2014, 'Mechanisms of CFTR Functional Variants That Impair Regulated Bicarbonate Permeation and Increase Risk for Pancreatitis but Not for Cystic Fibrosis', PLoS Genetics, vol. 10, no. 7, e1004376. https://doi.org/10.1371/journal.pgen.1004376

Mechanisms of CFTR Functional Variants That Impair Regulated Bicarbonate Permeation and Increase Risk for Pancreatitis but Not for Cystic Fibrosis. / LaRusch, Jessica; Jung, Jinsei; General, Ignacio J.; Lewis, Michele D.; Park, Hyun Woo; Brand, Randall E.; Gelrud, Andres; Anderson, Michelle A.; Banks, Peter A.; Conwell, Darwin; Lawrence, Christopher; Romagnuolo, Joseph; Baillie, John; Alkaade, Samer; Cote, Gregory; Gardner, Timothy B.; Amann, Stephen T.; Slivka, Adam; Sandhu, Bimaljit; Aloe, Amy; Kienholz, Michelle L.; Yadav, Dhiraj; Barmada, M. Michael; Bahar, Ivet; Lee, Min Goo; Whitcomb, David C.

In: PLoS Genetics, Vol. 10, No. 7, e1004376, 01.01.2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mechanisms of CFTR Functional Variants That Impair Regulated Bicarbonate Permeation and Increase Risk for Pancreatitis but Not for Cystic Fibrosis

AU - LaRusch, Jessica

AU - Jung, Jinsei

AU - General, Ignacio J.

AU - Lewis, Michele D.

AU - Park, Hyun Woo

AU - Brand, Randall E.

AU - Gelrud, Andres

AU - Anderson, Michelle A.

AU - Banks, Peter A.

AU - Conwell, Darwin

AU - Lawrence, Christopher

AU - Romagnuolo, Joseph

AU - Baillie, John

AU - Alkaade, Samer

AU - Cote, Gregory

AU - Gardner, Timothy B.

AU - Amann, Stephen T.

AU - Slivka, Adam

AU - Sandhu, Bimaljit

AU - Aloe, Amy

AU - Kienholz, Michelle L.

AU - Yadav, Dhiraj

AU - Barmada, M. Michael

AU - Bahar, Ivet

AU - Lee, Min Goo

AU - Whitcomb, David C.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev) cause complete loss of CFTR function and result in cystic fibrosis (CF), a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize that those CFTR mutations that disrupt the WNK1-SPAK activation mechanisms cause a selective, bicarbonate defect in channel function (CFTRBD) affecting organs that utilize CFTR for bicarbonate secretion (e.g. the pancreas, nasal sinus, vas deferens) but do not cause typical CF. To understand the structural and functional requirements of the CFTR bicarbonate-preferring channel, we (a) screened 984 well-phenotyped pancreatitis cases for candidate CFTRBD mutations from among 81 previously described CFTR variants; (b) conducted electrophysiology studies on clones of variants found in pancreatitis but not CF; (c) computationally constructed a new, complete structural model of CFTR for molecular dynamics simulation of wild-type and mutant variants; and (d) tested the newly defined CFTRBD variants for disease in non-pancreas organs utilizing CFTR for bicarbonate secretion. Nine variants (CFTR R74Q, R75Q, R117H, R170H, L967S, L997F, D1152H, S1235R, and D1270N) not associated with typical CF were associated with pancreatitis (OR 1.5, p = 0.002). Clones expressed in HEK 293T cells had normal chloride but not bicarbonate permeability and conductance with WNK1-SPAK activation. Molecular dynamics simulations suggest physical restriction of the CFTR channel and altered dynamic channel regulation. Comparing pancreatitis patients and controls, CFTRBD increased risk for rhinosinusitis (OR 2.3, p<0.005) and male infertility (OR 395, p≪0.0001). WNK1-SPAK pathway-activated increases in CFTR bicarbonate permeability are altered by CFTRBD variants through multiple mechanisms. CFTRBD variants are associated with clinically significant disorders of the pancreas, sinuses, and male reproductive system.

AB - CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev) cause complete loss of CFTR function and result in cystic fibrosis (CF), a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize that those CFTR mutations that disrupt the WNK1-SPAK activation mechanisms cause a selective, bicarbonate defect in channel function (CFTRBD) affecting organs that utilize CFTR for bicarbonate secretion (e.g. the pancreas, nasal sinus, vas deferens) but do not cause typical CF. To understand the structural and functional requirements of the CFTR bicarbonate-preferring channel, we (a) screened 984 well-phenotyped pancreatitis cases for candidate CFTRBD mutations from among 81 previously described CFTR variants; (b) conducted electrophysiology studies on clones of variants found in pancreatitis but not CF; (c) computationally constructed a new, complete structural model of CFTR for molecular dynamics simulation of wild-type and mutant variants; and (d) tested the newly defined CFTRBD variants for disease in non-pancreas organs utilizing CFTR for bicarbonate secretion. Nine variants (CFTR R74Q, R75Q, R117H, R170H, L967S, L997F, D1152H, S1235R, and D1270N) not associated with typical CF were associated with pancreatitis (OR 1.5, p = 0.002). Clones expressed in HEK 293T cells had normal chloride but not bicarbonate permeability and conductance with WNK1-SPAK activation. Molecular dynamics simulations suggest physical restriction of the CFTR channel and altered dynamic channel regulation. Comparing pancreatitis patients and controls, CFTRBD increased risk for rhinosinusitis (OR 2.3, p<0.005) and male infertility (OR 395, p≪0.0001). WNK1-SPAK pathway-activated increases in CFTR bicarbonate permeability are altered by CFTRBD variants through multiple mechanisms. CFTRBD variants are associated with clinically significant disorders of the pancreas, sinuses, and male reproductive system.

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