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Kidney Week

Abstract: TH-PO569

Unlocking the Role of Transcription Factors in PLCG2 Expression: Implications for Nephrotic Syndrome (NS)

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

  • Srivastava, Tarak, Children's Mercy Kansas City, Kansas City, Missouri, United States
  • Heruth, Daniel P., Children's Mercy Kansas City, Kansas City, Missouri, United States
  • Eddy, Sean, University of Michigan, Ann Arbor, Michigan, United States
  • Sharma, Mukut, Kansas City VA Medical Center, Kansas City, Missouri, United States
Background

The mechanism of immunopathogenesis of NS in minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS) remains unclear. We recently showed elevated expression of a scaffold protein SH3BP2 in MCD and FSGS. SH3BP2 forms a signaling complex with PLCG2, a key enzyme of immune signaling (JCI Insight 2024:e170055) and a candidate gene locus in NS (J Am Soc Nephrol. 2015:1701). To define the role of the SH3BP2/PLCG2 complex in NS we investigated the transcription factors (TFs) regulating PLCG2 expression.

Methods

RNA sequencing data (NEPTUNE consortium) from the glomerular compartment of Control (n=8), biopsy-proven MCD (n=89) and FSGS (n=93) cases were analyzed to (a) identify PLCG2 transcripts and (b) generate a Z-score for each gene for each patient. We used three models to predict TFs regulating PLCG2 expression: JASPAR (jaspar.elixir.no), TFBSpred (michalopoulos.net), and GeneCards (genecards.org). Gene Ontology (GO) analysis was performed using the Panther Knowledgebase (pantherdb.org).

Results

The primary transcript for PLCG2 (ENST00000564138.6; GRCh38.p14) was the most abundant. JASPAR, TFBSpred, and GeneCards predicted 665, 41, and 76 TFs regulating PLCG2, respectively. Data for the 56 TFs identified in more than one dataset were further analyzed. Gene expression for transcription factors IKZF1, MLX, IRF8, POU2F2 and SPI1 increased from Control to MCD to FSGS. Expression of MAX, MEIS2, GLIS2, PKNOX1, ETV2, TFE3, ELF4, SNAI2, ERG, ATF2, NR2C1, KLF9, KLF15, ATF7 and MAZ was significantly different in MCD and FSGS compared to Control. Expression for ETS1, SP1, TFDP1 and MITF was only significantly changed in MCD and that of SMAD5 and GABPA only in FSGS (p<0.05 in all comparisons). GO analysis of the 26 transcription factors identified key biological processes including: Developmental (20/26; FDR 8.87E-05), Immune System (11/26; 1.14E-02), Positive Regulation Transcription by RNA PolII (20/26; 6.25E-16) and Negative Regulation of Transcription by RNA PolII (13/26; 2.07E-03).

Conclusion

Identification of these 26 transcriptional regulators of PLCG2 that play a role in Immune System, Regulation of Transcription by RNA PolII and Developmental processes will provide the basis for investigating the role of PLCG2/SH3BP2 complex in the immunopathogenesis of NS and for identifying therapeutic targets.

Funding

  • Private Foundation Support