cAMP-mediated regulation of cholesterol accumulation in cystic fibrosis and Niemann-Pick type C cells.
The goal of this study was to identify a mechanism regulating cholesterol accumulation in cystic fibrosis (CF) cells. Both CFTR activation and expression are regulated by the cAMP pathway, and it is hypothesized that a feedback response involving this pathway may be involved in the phenotype of cholesterol accumulation. To examine the role of the cAMP pathway in cholesterol accumulation, we treated two CF model cell lines with the Rp diastereomer of adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS) and visualized by filipin staining. Rp-cAMPS treatment eliminated cholesterol accumulation in CF cells, whereas 8-bromo-cAMP treatment led to cholesterol accumulation in wild-type cells. To confirm these findings in an independent model system, we also examined the role of cAMP in modulating cholesterol accumulation in Niemann-Pick type C (NPC) fibroblasts. Expression of the protein related to NPC, NPC1, is also directly regulated by cAMP; therefore, it is postulated that NPC cells exhibit the same cAMP-mediated control of cholesterol accumulation. Cholesterol accumulation in NPC cells also was reduced by the presence of Rp-cAMPS. Expression of beta-arrestin-2 (betaarr2), a marker of cellular response to cAMP signaling, was significantly elevated in CF model cells, Cftr(-/-) MNE, primary tissue obtained by nasal scrapes from CF subjects, and in NPC fibroblasts compared with respective controls.
Manson
Mary E
ME
Department of Pediatrics, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH 44106-4948, USA.
Corey
Deborah A
DA
White
Nicole M
NM
Kelley
Thomas J
TJ
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D019390:Arrestins / Q000378:metabolism; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology*; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D051379:Mice; D052556:Niemann-Pick Disease, Type C / Q000378:metabolism* / Q000473:pathology*; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D013873:Thionucleotides / Q000494:pharmacology; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
C000605254:ARRB2 protein, human; C000605258:Arrb2 protein, mouse; D019390:Arrestins; D017362:Cyclic AMP Response Element-Binding Protein; D018343:Receptors, Adrenergic, beta-2; D013873:Thionucleotides; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D015124:8-Bromo Cyclic Adenosine Monophosphate; C016957:adenosine-3',5'-cyclic phosphorothioate; D002784:Cholesterol; D000242:Cyclic AMP
Clin Sci (Lond). 1990 Apr;78(4):409-17
1971204
J Pathol. 2000 Feb;190(2):126-32
10657009
Mol Endocrinol. 2003 Apr;17(4):704-15
12554781
Am J Physiol Lung Cell Mol Physiol. 2007 Feb;292(2):L476-86
17085523
J Biol Chem. 2003 Nov 7;278(45):44238-45
12944399
Arch Biochem Biophys. 2002 Oct 1;406(1):33-9
12234487
J Cell Sci. 2007 Feb 1;120(Pt 3):447-55
17213331
Am J Respir Crit Care Med. 1997 Oct;156(4 Pt 1):1197-204
9351622
Int J Biochem Cell Biol. 2008;40(3):432-46
17936667
Pediatr Pulmonol. 2005 Dec;40(6):500-10
16208679
J Immunol. 2000 Apr 1;164(7):3870-7
10725749
Eur J Pharmacol. 2002 Feb 2;436(3):165-72
11858796
Am J Physiol Lung Cell Mol Physiol. 2007 Feb;292(2):L383-95
16920886
Am J Physiol Lung Cell Mol Physiol. 2003 Dec;285(6):L1286-95
12948935
Am J Physiol Lung Cell Mol Physiol. 2001 Jul;281(1):L71-8
11404248
Nat Med. 2008 Apr;14(4):382-91
18376404
Am J Physiol Lung Cell Mol Physiol. 2004 Nov;287(5):L944-52
15246977
Am J Physiol. 1995 Nov;269(5 Pt 1):L625-30
7491981
Am J Respir Cell Mol Biol. 2000 Aug;23(2):121-7
10919974
Am J Physiol Lung Cell Mol Physiol. 2007 Nov;293(5):L1250-60
17827250
Am J Physiol Lung Cell Mol Physiol. 2001 Mar;280(3):L493-502
11159033
Am J Physiol. 1986 Oct;251(4 Pt 2):R818-22
3021002
J Interferon Cytokine Res. 2006 Sep;26(9):609-27
16978065
FASEB J. 2008 Aug;22(8):2872-9
18424767
Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7500-4
1715567
J Biol Chem. 2005 Mar 4;280(9):7997-8003
15611099
Proc Natl Acad Sci U S A. 2004 May 18;101(20):7693-8
15136726
Am J Respir Crit Care Med. 2004 Mar 1;169(5):645-53
14670800
Am J Physiol. 1996 Jul;271(1 Pt 1):L85-92
8760136
Hum Gene Ther. 2004 Dec;15(12):1255-69
15684701
J Clin Invest. 1998 Jun 1;101(11):2598-605
9616231
Life Sci. 1998;62(17-18):1561-5
9585136
J Clin Invest. 1997 Dec 1;100(11):2810-5
9389746
J Biol Chem. 2004 Nov 12;279(46):48214-23
15347664
Am J Physiol Lung Cell Mol Physiol. 2004 Aug;287(2):L374-81
15107292
J Cell Sci. 1999 Feb;112 ( Pt 3):329-38
9885286
Am J Respir Cell Mol Biol. 2004 Nov;31(5):538-43
15256387
EMBO Rep. 2006 May;7(5):553-9
16612392
J Clin Invest. 2000 Aug;106(3):403-10
10930443
J Biol Chem. 1996 Dec 13;271(50):31869-77
8943230
J Pathol. 1998 Mar;184(3):323-31
9614386
J Biol Chem. 1986 Dec 15;261(35):16769-74
3782141
J Pathol. 2007 Feb;211(3):340-50
17186573
J Clin Invest. 1998 Sep 15;102(6):1200-7
9739054
Eur Arch Otorhinolaryngol. 2002 Apr;259(4):222-6
12064512