Limits...
CARD14 expression in dermal endothelial cells in psoriasis.

Harden JL, Lewis SM, Pierson KC, Suárez-Fariñas M, Lentini T, Ortenzio FS, Zaba LC, Goldbach-Mansky R, Bowcock AM, Lowes MA - PLoS ONE (2014)

Bottom Line: CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling.CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells.Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, United States of America.

ABSTRACT
Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.

Show MeSH

Related in: MedlinePlus

Increased expression of CARD14-modulated chemokines in psoriatic skin co-localized with dermal endothelial cells.(a) Immunohistochemistry of CXCL1, CCL2, and CCL5 in non-lesional and lesional skin of both classical psoriasis (left, representative images) and patient GEN001 (right). (b) Two-color immunofluorescence of these chemokines (red) and CD31+ endothelial cells (green) in classical psoriasis lesional skin. Representative images; bar = 10 µm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4219711&req=5

pone-0111255-g005: Increased expression of CARD14-modulated chemokines in psoriatic skin co-localized with dermal endothelial cells.(a) Immunohistochemistry of CXCL1, CCL2, and CCL5 in non-lesional and lesional skin of both classical psoriasis (left, representative images) and patient GEN001 (right). (b) Two-color immunofluorescence of these chemokines (red) and CD31+ endothelial cells (green) in classical psoriasis lesional skin. Representative images; bar = 10 µm.

Mentions: Several of the chemokines determined to be upregulated in ECs via mutant CARD14 have been previously identified as significantly upregulated genes and proteins in psoriasis, on several platforms [16]. To determine if our in vitro HDBEC CARD14 transfection findings could be supported in vivo, skin biopsies were stained for chemokines by immunohistochemistry. Indeed, LS skin contained significantly more CCL2, CCL5, and CXCL1 compared to NL skin, in both classical psoriasis and the GEN001 patient (Figure 5a). Two-color immunofluorescence studies confirmed that dermal CXCL1, CCL2, and CCL5 co-localized with ECs (Figure 5b). Additionally, HDBECs cultured with IL-17, TNFα, and IFNγ, three cytokines heavily implicated in psoriasis pathogenesis, resulted in upregulation of IL-8, CXCL1, CXCL10, and CCL5 (Figure S9 in File S1). In conclusion, chemokines that were upregulated by transfection of psoriasis-associated CARD14 mutations were found both endogenously in dermal EC within psoriatic lesional skin, as well as upregulated in dermal ECs exposed to key psoriatic cytokines. This suggests a connection between CARD14-NF-κB activity in endothelial cells and subsequent chemokine induction in psoriasis, and a potential role for CARD14 activity within endothelial cells in the pathogenesis of psoriasis.


CARD14 expression in dermal endothelial cells in psoriasis.

Harden JL, Lewis SM, Pierson KC, Suárez-Fariñas M, Lentini T, Ortenzio FS, Zaba LC, Goldbach-Mansky R, Bowcock AM, Lowes MA - PLoS ONE (2014)

Increased expression of CARD14-modulated chemokines in psoriatic skin co-localized with dermal endothelial cells.(a) Immunohistochemistry of CXCL1, CCL2, and CCL5 in non-lesional and lesional skin of both classical psoriasis (left, representative images) and patient GEN001 (right). (b) Two-color immunofluorescence of these chemokines (red) and CD31+ endothelial cells (green) in classical psoriasis lesional skin. Representative images; bar = 10 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4219711&req=5

pone-0111255-g005: Increased expression of CARD14-modulated chemokines in psoriatic skin co-localized with dermal endothelial cells.(a) Immunohistochemistry of CXCL1, CCL2, and CCL5 in non-lesional and lesional skin of both classical psoriasis (left, representative images) and patient GEN001 (right). (b) Two-color immunofluorescence of these chemokines (red) and CD31+ endothelial cells (green) in classical psoriasis lesional skin. Representative images; bar = 10 µm.
Mentions: Several of the chemokines determined to be upregulated in ECs via mutant CARD14 have been previously identified as significantly upregulated genes and proteins in psoriasis, on several platforms [16]. To determine if our in vitro HDBEC CARD14 transfection findings could be supported in vivo, skin biopsies were stained for chemokines by immunohistochemistry. Indeed, LS skin contained significantly more CCL2, CCL5, and CXCL1 compared to NL skin, in both classical psoriasis and the GEN001 patient (Figure 5a). Two-color immunofluorescence studies confirmed that dermal CXCL1, CCL2, and CCL5 co-localized with ECs (Figure 5b). Additionally, HDBECs cultured with IL-17, TNFα, and IFNγ, three cytokines heavily implicated in psoriasis pathogenesis, resulted in upregulation of IL-8, CXCL1, CXCL10, and CCL5 (Figure S9 in File S1). In conclusion, chemokines that were upregulated by transfection of psoriasis-associated CARD14 mutations were found both endogenously in dermal EC within psoriatic lesional skin, as well as upregulated in dermal ECs exposed to key psoriatic cytokines. This suggests a connection between CARD14-NF-κB activity in endothelial cells and subsequent chemokine induction in psoriasis, and a potential role for CARD14 activity within endothelial cells in the pathogenesis of psoriasis.

Bottom Line: CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling.CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells.Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, United States of America.

ABSTRACT
Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.

Show MeSH
Related in: MedlinePlus