The methylation of the TSC2 promoter underlies the abnormal growth of TSC2 angiomyolipoma-derived smooth muscle cells

Elena Lesma, Silvia Maria Sirchia, Silvia Ancona, Stephana Carelli, Silvano Bosari, Filippo Ghelma, Emanuele Montanari, Anna Maria Di Giulio, Alfredo Gorio

Research output: Contribution to journalArticle

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Abstract

Tuberous sclerosis complex (TSC) is an autosomal-dominant disease that is caused by mutations in either the TSC1 or TSC2 gene. Smooth muscle-like cells (ASMs) were isolated from an angiomyolipoma of a patient with TSC. These cells lacked tuberin, were labeled by both HMB45 and CD44v6 antibodies, and had constitutive S6 phosphorylation. The cells bear a germline TSC2 intron 8-exon 9 junction mutation, but DNA analysis and polymerase chain reaction amplification failed to demonstrate loss of heterozygosity. Testing for an epigenetic alteration, we detected methylation of the TSC2 promoter. Its biological relevance was confirmed by tuberin expression and a reduction in HMB45 labeling and S6 constitutive phosphorylation after exposure to the chromatin-remodeling agents, trichostatin A and 5-azacytidine. These cells were named TSC2 -/meth ASMs. Their proliferation required epidermal growth factor in the medium as previously described for TSC2-/- ASMs. Blockade of epidermal growth factor with monoclonal antibodies caused the death of TSC2 -/meth ASMs. In addition, rapamycin effectively blocked the proliferation of these cells. Our data show for the first time that methylation of the TSC2 promoter might cause a complete loss of tuberin in TSC2 cells , and that the pathogenesis of angiomyolipomas might also originate from epigenetic defects in smooth muscle cells. Additionally, the effect of chromatin-remodeling agents in these cells suggests a further avenue for the treatment of TSC as well as lymphangioleiomyomatosis.

Original languageEnglish
Pages (from-to)2150-2159
Number of pages10
JournalAmerican Journal of Pathology
Volume174
Issue number6
DOIs
Publication statusPublished - Jun 2009

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Angiomyolipoma
Methylation
Smooth Muscle Myocytes
Tuberous Sclerosis
S 6
Growth
Chromatin Assembly and Disassembly
Epidermal Growth Factor
Epigenomics
trichostatin A
Phosphorylation
Lymphangioleiomyomatosis
Azacitidine
Mutation
Loss of Heterozygosity
Sirolimus
DNA-Directed DNA Polymerase
Introns
Exons
Monoclonal Antibodies

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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The methylation of the TSC2 promoter underlies the abnormal growth of TSC2 angiomyolipoma-derived smooth muscle cells. / Lesma, Elena; Sirchia, Silvia Maria; Ancona, Silvia; Carelli, Stephana; Bosari, Silvano; Ghelma, Filippo; Montanari, Emanuele; Di Giulio, Anna Maria; Gorio, Alfredo.

In: American Journal of Pathology, Vol. 174, No. 6, 06.2009, p. 2150-2159.

Research output: Contribution to journalArticle

Lesma, E, Sirchia, SM, Ancona, S, Carelli, S, Bosari, S, Ghelma, F, Montanari, E, Di Giulio, AM & Gorio, A 2009, 'The methylation of the TSC2 promoter underlies the abnormal growth of TSC2 angiomyolipoma-derived smooth muscle cells', American Journal of Pathology, vol. 174, no. 6, pp. 2150-2159. https://doi.org/10.2353/ajpath.2009.080799
Lesma, Elena ; Sirchia, Silvia Maria ; Ancona, Silvia ; Carelli, Stephana ; Bosari, Silvano ; Ghelma, Filippo ; Montanari, Emanuele ; Di Giulio, Anna Maria ; Gorio, Alfredo. / The methylation of the TSC2 promoter underlies the abnormal growth of TSC2 angiomyolipoma-derived smooth muscle cells. In: American Journal of Pathology. 2009 ; Vol. 174, No. 6. pp. 2150-2159.
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AU - Bosari, Silvano

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AB - Tuberous sclerosis complex (TSC) is an autosomal-dominant disease that is caused by mutations in either the TSC1 or TSC2 gene. Smooth muscle-like cells (ASMs) were isolated from an angiomyolipoma of a patient with TSC. These cells lacked tuberin, were labeled by both HMB45 and CD44v6 antibodies, and had constitutive S6 phosphorylation. The cells bear a germline TSC2 intron 8-exon 9 junction mutation, but DNA analysis and polymerase chain reaction amplification failed to demonstrate loss of heterozygosity. Testing for an epigenetic alteration, we detected methylation of the TSC2 promoter. Its biological relevance was confirmed by tuberin expression and a reduction in HMB45 labeling and S6 constitutive phosphorylation after exposure to the chromatin-remodeling agents, trichostatin A and 5-azacytidine. These cells were named TSC2 -/meth ASMs. Their proliferation required epidermal growth factor in the medium as previously described for TSC2-/- ASMs. Blockade of epidermal growth factor with monoclonal antibodies caused the death of TSC2 -/meth ASMs. In addition, rapamycin effectively blocked the proliferation of these cells. Our data show for the first time that methylation of the TSC2 promoter might cause a complete loss of tuberin in TSC2 cells , and that the pathogenesis of angiomyolipomas might also originate from epigenetic defects in smooth muscle cells. Additionally, the effect of chromatin-remodeling agents in these cells suggests a further avenue for the treatment of TSC as well as lymphangioleiomyomatosis.

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