Lupus Nephritis IgG Induction of Calcium/Calmodulin‐Dependent Protein Kinase IV Expression in Podocytes and Alteration of Their Function

• Published in: Arthritis & rheumatology (Hoboken, N.J.) Vol. 68; no. 4; pp. 944 - 952
• Main Authors: Ichinose, Kunihiro, Ushigusa, Takeshi, Nishino, Ayako, Nakashima, Yosikazu, Suzuki, Takahisa, Horai, Yoshiro, Koga, Tomohiro, Kawashiri, Shin‐ya, Iwamoto, Naoki, Tamai, Mami, Arima, Kazuhiko, Nakamura, Hideki, Obata, Yoko, Yamamoto, Kazuo, Origuchi, Tomoki, Nishino, Tomoya, Kawakami, Atsushi, Tsokos, George C.
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.04.2016
• Subjects: Adult; Animals; B7-2 Antigen - drug effects; B7-2 Antigen - genetics; B7-2 Antigen - metabolism; Blotting, Western; Calcium; Calcium-Calmodulin-Dependent Protein Kinase Type 4 - drug effects; Calcium-Calmodulin-Dependent Protein Kinase Type 4 - genetics; Calcium-Calmodulin-Dependent Protein Kinase Type 4 - metabolism; Female; Fluorescent Antibody Technique; Gene expression; Gene Expression - drug effects; Gene Expression Profiling; Gene Knockdown Techniques; Histocompatibility Antigens Class I - genetics; Histocompatibility Antigens Class I - metabolism; Humans; Immunoglobulin G - pharmacology; In Situ Hybridization; In Vitro Techniques; Kidney Glomerulus - metabolism; Kinases; Lupus; Lupus Nephritis - immunology; Lupus Nephritis - metabolism; Membrane Proteins - metabolism; Mice; Mice, Inbred MRL lpr; Mice, Knockout; Middle Aged; Oligonucleotide Array Sequence Analysis; Podocytes - drug effects; Podocytes - metabolism; Proteins; Receptors, Fc - genetics; Receptors, Fc - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Up-Regulation - drug effects; Young Adult
• ISSN: 2326-5191; 2326-5205
• DOI: 10.1002/art.39499

Objective Kidney podocytes and their slit diaphragms prevent urinary protein loss. T cells from patients with systemic lupus erythematosus display increased expression of calcium/calmodulin‐dependent protein kinase IV (CaMKIV). The present study was undertaken to investigate the role of CaMKIV in podocyte function in lupus nephritis (LN). Methods We treated kidney podocytes with IgG derived from healthy individuals or patients with LN and then analyzed gene expression using a DNA microarray. The localization of IgG in podocytes was analyzed by immunofluorescence staining, with or without silencing of neonatal Fc receptor (FcRn). In addition, we silenced CAMK4 in podocytes and analyzed the expression of selected genes. We also examined the expression of CD86 in kidney podocytes from MRL/lpr, MRL/lpr.camkiv–/–, and MRL/MPJ mice by in situ hybridization. Results We found that exposure of podocytes to IgG resulted in entry of IgG into the cytoplasm. IgG entered podocytes via the FcRn because less IgG was found in the cytoplasm of podocytes treated with FcRn small interfering RNA. DNA microarray studies of podocytes exposed to LN‐derived IgG revealed up‐regulation of genes related to the activation of immune cells or podocyte damage. Interestingly, CD86 expression decreased after silencing CAMK4 in podocytes. Also, in situ hybridization experiments showed that the expression of CD86 was reduced in podocytes from MRL/lpr.camkiv–/– mice. Conclusion LN‐derived IgG enters podocytes and up‐regulates CAMK4, which is followed by increased expression of genes known to be linked to podocyte damage and T cell activation. Targeted inhibition of CAMK4 in podocytes may prove to be clinically useful in patients with LN.