Recognition of tuberous sclerosis in adult women: delayed presentation with life-threatening consequences.

BACKGROUND: Tuberous sclerosis complex (TSC) is associated with tumor development in the brain, retina, kidney, skin, heart, and lung. Seizures, intellectual disability, and characteristic skin lesions commonly manifest in early childhood, but some findings, notably renal angiomyolipomas and pulmonary lymphangioleiomyomatosis (LAM), emerge later, placing adults with undiagnosed TSC at increased risk for morbidity and mortality. OBJECTIVE: To describe the clinical presentation and severity of TSC in adult women. DESIGN: Retrospective cohort study. SETTING: National Institutes of Health Clinical Center, Bethesda, Maryland, 1995 to 2010. PATIENTS: 79 women aged 18 years or older who were enrolled in an observational cohort study of TSC to evaluate disease manifestations. MEASUREMENTS: History, physical examination, pulmonary function testing, chest radiography, abdominal computed tomography, high-resolution chest computed tomography, and brain magnetic resonance imaging were used to evaluate patients. RESULTS: Among the 45 patients who received a diagnosis of TSC in adulthood, 21 presented with symptoms due to LAM, 19 with renal angiomyolipomas, and 10 with seizures. Of the 45 patients, 30 met clinical criteria for TSC in childhood that remained undiagnosed for a median of 21.5 years and 15 were older than 18 years before meeting the clinical criteria for TSC. Patients diagnosed in adulthood and those diagnosed in childhood had similar occurrences of pneumothorax, shortness of breath, hemoptysis, nephrectomy, and death. LIMITATION: No men were included in the study, and selection was biased toward patients having pulmonary LAM. CONCLUSION: Women who received a TSC diagnosis in adulthood had minimal morbidity during childhood but were still at risk for life-threatening pulmonary and renal manifestations. PRIMARY FUNDING SOURCE: Intramural Research Program, National Institutes of Health, National Heart, Lung, and Blood Institute.

MCP-1 overexpressed in tuberous sclerosis lesions acts as a paracrine factor for tumor development.

Patients with tuberous sclerosis complex (TSC) develop hamartomatous tumors showing loss of function of the tumor suppressor TSC1 (hamartin) or TSC2 (tuberin) and increased angiogenesis, fibrosis, and abundant mononuclear phagocytes. To identify soluble factors with potential roles in TSC tumorigenesis, we screened TSC skin tumor-derived cells for altered gene and protein expression. Fibroblast-like cells from 10 angiofibromas and five periungual fibromas produced higher levels of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein than did fibroblasts from the same patient's normal skin. Conditioned medium from angiofibroma cells stimulated chemotaxis of a human monocytic cell line to a greater extent than conditioned medium from TSC fibroblasts, an effect blocked by neutralizing MCP-1-specific antibody. Overexpression of MCP-1 seems to be caused by loss of tuberin function because Eker rat embryonic fibroblasts null for Tsc2 (EEF Tsc2(-/-)) produced 28 times as much MCP-1 protein as did EEF Tsc2(+/+) cells; transient expression of WT but not mutant human TSC2 by EEF Tsc2(-/-) cells inhibited MCP-1 production; and pharmacological inhibition of the Rheb-mTOR pathway, which is hyperactivated after loss of TSC2, decreased MCP-1 production by EEF Tsc2(-/-) cells. Together these findings suggest that MCP-1 is an important paracrine factor for TSC tumorigenesis and may be a new therapeutic target.

Mesenchymal-epithelial interactions involving epiregulin in tuberous sclerosis complex hamartomas.

Patients with tuberous sclerosis complex (TSC) develop hamartomas containing biallelic inactivating mutations in either TSC1 or TSC2, resulting in mammalian target of rapamycin (mTOR) activation. Hamartomas overgrow epithelial and mesenchymal cells in TSC skin. The pathogenetic mechanisms for these changes had not been investigated, and the existence or location of cells with biallelic mutations ("two-hit" cells) was unclear. We compared TSC skin hamartomas (angiofibromas and periungual fibromas) with normal-appearing skin of the same patient, and we observed more proliferation and mTOR activation in hamartoma epidermis. Two-hit cells were not detected in the epidermis. Fibroblast-like cells in the dermis, however, exhibited allelic deletion of TSC2, in both touch preparations of fresh tumor samples and cells grown from TSC skin tumors, suggesting that increased epidermal proliferation and mTOR activation were not caused by second-hit mutations in the keratinocytes but by mesenchymal-epithelial interactions. Gene expression arrays, used to identify potential paracrine factors released by mesenchymal cells, revealed more epiregulin mRNA in fibroblast-like angiofibroma and periungual fibroma cells than in fibroblasts from normal-appearing skin of the same patient. Elevation of epiregulin mRNA was confirmed with real-time PCR, and increased amounts of epiregulin protein were demonstrated with immunoprecipitation. Epiregulin stimulated keratinocyte proliferation and phosphorylation of ribosomal protein S6 in vitro. These results suggest that hamartomatous TSC skin tumors are induced by paracrine factors released by two-hit cells in the dermis and that proliferation with mTOR activation of the overlying epidermis is an effect of epiregulin.

Cutaneous endothelial cell activation in normal skin of patients with dermatitis herpetiformis associated with increased serum levels of IL-8, sE-Selectin, and TNF-alpha.

The mechanisms that lead to the development of skin lesions in patients with dermatitis herpetiformis (DH) are not known. We hypothesized that an ongoing immune response in the gut of patients with DH would result in an increase in circulating cytokines and be associated with endothelial cell activation, creating a proinflammatory environment in the skin. Skin biopsies from the normal-appearing inner arm of 11 DH patients, with no active skin lesions, and 12 normal subjects were analyzed for E-selectin (E-sel) and ICAM-1 mRNA. DH patients' skin expressed markedly increased levels of E-sel mRNA. Mean E-sel mRNA expression in DH skin was 1,271 (range 63.78-5861) times greater than that of a control, normal skin (P<0.001) with no significant increased expression of ICAM-1 mRNA. Serum levels of soluble E-selectin (sE-sel), IgA anti-tissue transglutaminase antibodies, and serum IL-8 levels were significantly increased in patients with DH. These studies demonstrate that patients with DH have evidence of endothelial cell activation in the skin and systemic manifestations of the ongoing inflammation associated with the mucosal immune response. Endothelial cell activation may play a critical role in the development of skin lesions in patients with DH and may represent a common mechanism for cutaneous manifestations of inflammatory gastrointestinal diseases.

Serum IL-8 in patients with dermatitis herpetiformis is produced in response to dietary gluten.

Patients with dermatitis herpetiformis (DH) have a gluten-sensitive enteropathy and while on gluten-containing diets have elevated levels of serum IL-8. We hypothesized that the mucosal immune response to gluten is responsible for the elevated serum IL-8. Six DH patients were studied while on a gluten-free diet (GFD), whereas four continued on a normal diet. Patients were followed for a mean 2.2 years and serum IL-8 was analyzed. Small bowel biopsies from five DH patients on normal diets, two DH patients on GFD, and six subjects with no small bowel abnormalities were analyzed for IL-8 mRNA. Serum IL-8 levels normalized in five of six patients on GFD and decreased in one, whereas serum IL-8 levels showed no statistically significant change in DH patients on normal diets. Small bowel biopsies from DH patients on normal diets had increased expression of IL-8 mRNA compared to normal subjects, whereas patients on a GFD showed no significant increase in small bowel mRNA. No significant IL-8 mRNA was detected in normal skin biopsies from patients with DH. These observations suggest that the IL-8 in the serum of patients with DH originates from the small bowel as a mucosal immune response to gluten ingestion.

Increased E-selectin, IL-8 and IL-10 gene expression in human skin after minimal trauma.

While clinical observations suggest that trauma to the skin plays a critical role in the induction of skin lesions in some skin diseases, the mechanism by which these lesions are induced is not known. We have postulated that minor trauma to the skin may lead to the expression of critical adhesion molecules on epidermal endothelial cells (E-selectin) and pro-inflammatory cytokines, which would predispose these areas to the development of skin lesions. In order to test this hypothesis normal inner arm skin of 11 normal subjects was gently rubbed with a pencil eraser for 2 min. Four hours after rubbing, skin biopsies were obtained from the rubbed site and from adjacent normal, unrubbed inner arm skin. Expression of E-selectin, intercellular adhesion molecules (ICAM-1) and the mRNA of selected cytokines was studied utilizing real time polymerase chain reactions. Biopsies were also examined for the presence of an inflammatory infiltrate and for the presence of E-selectin and ICAM-1. No clinical or histologic changes were seen in the skin expression/unrubbed skin expression = 9.0; (median ratio rubbed skin expression/unrubbed skin expression range 0.9-161.0), ICAM-1 (median rubbed skin expression/unrubbed skin expression = 3.2; range 0.9-19.8), IL-8 (median rubbed skin expression/unrubbed skin expression = 6.6; range 2.6-57.3) and IL-10 (median rubbed skin expression/unrubbed skin expression = 13.1; range 2.4-29.0) was noted. Immunohistochemistry revealed the presence of E-selectin in the dermal blood vessels in three of four subjects 4 h after rubbing but not in the unrubbed skin. Changes in ICAM -1 or HLA-DR deposits were seen in the rubbed compared with the unrubbed skin. These findings demonstrated that minor trauma to skin may induce expression of E-selectin, ICAM-1 and IL-8, which may make the skin a more permissive site for the development of inflammatory reactions. These findings may play an important role in the development of skin lesions in areas of minor trauma.