High-Throughput Functional Analysis of CFTR and Other Apically Localized Proteins in iPSC-Derived Human Intestinal Organoids

被引：12

作者：

Xia, Sunny ^{[1
,2
,3
]}

Bozoky, Zoltan ^{[1
]}

Di Paola, Michelle ^{[3
]}

Laselva, Onofrio ^{[1
,4
]}

Ahmadi, Saumel ^{[5
]}

Jiang, Jia Xin ^{[1
]}

Pitstick, Amy L. ^{[6
]}

Jiang, Chong ^{[2
]}

Rotin, Daniela ^{[2
,7
]}

Mayhew, Christopher N.

Jones, Nicola L. ^{[2
,3
,8
]}

Bear, Christine E. ^{[1
,3
,7
]}

机构：

[1] Hosp Sick Children, Mol Med, 686 Bay St, Toronto, ON M5G 0A4, Canada

[2] Hosp Sick Children, Cell Biol, Toronto, ON M5G 0A4, Canada

[3] Univ Toronto, Dept Physiol, Toronto, ON M5S 1A8, Canada

[4] Univ Foggia, Dept Med & Surg Sci, I-71122 Foggia, Italy

[5] Washington Univ, Dept Neurol, St Louis, MO 63110 USA

[6] Cincinnati Childrens Hosp Med Ctr, Div Dev Biol, Cincinnati, OH 45229 USA

[7] Univ Toronto, Dept Biochem, Toronto, ON M5G 0A4, Canada

[8] Univ Toronto, Dept Paediat, Toronto, ON M5G 0A4, Canada

来源：

CELLS | 2021年 / 10卷 / 12期

关键词：

cystic fibrosis; high throughput; in vitro models; CFTR; ENaC; ion channel activity; CYSTIC-FIBROSIS; CELLS; CONDUCTANCE; THERAPY; ENAC;

D O I：

10.3390/cells10123419

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived organoids have only been used to study pharmacological modulation of mutant CFTR channel activity and not the activity of other disease-relevant membrane protein constituents. In the current work, we describe a high-throughput, fluorescence-based assay of CFTR channel activity in iPSC-derived intestinal organoids and describe how this method can be adapted to study other apical membrane proteins. Specifically, we show how this assay can be employed to study CFTR and ENaC channels and an electrogenic acid transporter in the same iPSC-derived intestinal tissue. This phenotypic platform promises to expand CF therapy discovery to include strategies that target multiple determinants of epithelial fluid transport.

引用

页数：17

共 32 条

[1] SLC6A14, an amino acid transporter, modifies the primary CF defect in fluid secretion [J].

Ahmadi, Saumel ;

Xia, Sunny ;

Wu, Yu-Sheng ;

Di Paola, Michelle ;

Kissoon, Randolph ;

Luk, Catherine ;

Lin, Fan ;

Du, Kai ;

Rommens, Johanna ;

Bear, Christine E. .

ELIFE, 2018, 7

[2] Phenotypic profiling of CFTR modulators in patient-derived respiratory epithelia [J].

Ahmadi, Saumel ;

Bozoky, Zoltan ;

Di Paola, Michelle ;

Xia, Sunny ;

Li, Canhui ;

Wong, Amy P. ;

Wellhauser, Leigh ;

Molinski, Steven V. ;

Ip, Wan ;

Ouyang, Hong ;

Avolio, Julie ;

Forman-Kay, Julie D. ;

Ratjen, Felix ;

Hirota, Jeremy A. ;

Rommens, Johanna ;

Rossant, Janet ;

Gonska, Tanja ;

Moraes, Theo J. ;

Bear, Christine E. .

NPJ GENOMIC MEDICINE, 2017, 2

[3] Mechanisms of ENaC regulation and clinical implications [J].

Bhalla, Vivek ;

Hallows, Kenneth R. .

JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2008, 19 (10) :1845-1854

[4] Validation and Optimization of Novel High-Throughput Assays for Human Epithelial Sodium Channels [J].

Chen, Mao Xiang ;

Gatfield, Kelly ;

Ward, Emma ;

Downie, David ;

Sneddon, Helen F. ;

Walsh, Stacey ;

Powell, Andrew J. ;

Laine, Dramane ;

Carr, Michael ;

Trezise, Derek .

JOURNAL OF BIOMOLECULAR SCREENING, 2015, 20 (02) :242-253

[5] Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis [J].

Dekkers, Johanna F. ;

Berkers, Gitte ;

Kruisselbrink, Evelien ;

Vonk, Annelotte ;

de Jonge, Hugo R. ;

Janssens, Hettie M. ;

Bronsveld, Inez ;

de Graaf, Eduard A. van ;

Nieuwenhuis, Edward E. S. ;

Houwen, Roderick H. J. ;

Vleggaar, Frank P. ;

Escher, Johanna C. ;

de Rijke, Yolanda B. ;

Majoor, Christof J. ;

Heijerman, Harry G. M. ;

de Winter-de Groot, Karin M. ;

Clevers, Hans ;

van der Ent, Cornelis K. ;

Beekman, Jeffrey M. .

SCIENCE TRANSLATIONAL MEDICINE, 2016, 8 (344)

[6] A functional CFTR assay using primary cystic fibrosis intestinal organoids [J].

Dekkers, Johanna F. ;

Wiegerinck, Caroline L. ;

de Jonge, Hugo R. ;

Bronsveld, Inez ;

Janssens, Hettie M. ;

de Winter-de Groot, Karin M. ;

Brandsma, Arianne M. ;

de Jong, Nienke W. M. ;

Bijvelds, Marcel J. C. ;

Scholte, Bob J. ;

Nieuwenhuis, Edward E. S. ;

van den Brink, Stieneke ;

Clevers, Hans ;

van der Ent, Cornelis K. ;

Middendorp, Sabine ;

Beekman, Jeffrey M. .

NATURE MEDICINE, 2013, 19 (07) :939-+

[7] SLC6A14 Is a Genetic Modifier of Cystic Fibrosis That Regulates Pseudomonas aeruginosa Attachment to Human Bronchial Epithelial Cells [J].

Di Paola, Michelle ;

Park, Amber J. ;

Ahmadi, Saumel ;

Roach, Elyse J. ;

Wu, Yu-Sheng ;

Struder-Kypke, Michaela ;

Lam, Joseph S. ;

Bear, Christine E. ;

Khursigara, Cezar M. .

MBIO, 2017, 8 (06)

[8] The CF Canada-Sick Kids Program in individual CF therapy: A resource for the advancement of personalized medicine in CF [J].

Eckford, Paul D. W. ;

McCormack, Jacqueline ;

Munsie, Lise ;

He, Gengming ;

Stanojevic, Sanja ;

Pereira, Sergio L. ;

Ho, Karen ;

Avolio, Julie ;

Bartlett, Claire ;

Yang, Jin Ye ;

Wong, Amy P. ;

Wellhauser, Leigh ;

Huan, Ling Jun ;

Jiang, Jia Xin ;

Ouyang, Hong ;

Du, Kai ;

Klingel, Michelle ;

Kyriakopoulou, Lianna ;

Gonska, Tanja ;

Moraes, Theo J. ;

Strug, Lisa J. ;

Rossant, Janet ;

Ratjen, Felix ;

Bear, Christine E. .

JOURNAL OF CYSTIC FIBROSIS, 2019, 18 (01) :35-43

[9] Evaluation of second generation amiloride analogs as therapy for cystic fibrosis lung disease [J].

Hirsh, AJ ;

Sabater, JR ;

Zamurs, A ;

Smith, RT ;

Paradiso, AM ;

Hopkins, S ;

Abraham, WM ;

Boucher, RC .

JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, 2004, 311 (03) :929-938

[10] Does epithelial sodium channel hyperactivity contribute to cystic fibrosis lung disease? [J].

Hobbs, Carey A. ;

Tan, Chong Da ;

Tarran, Robert .

JOURNAL OF PHYSIOLOGY-LONDON, 2013, 591 (18) :4377-4387

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