Program

PO1-3-11

Histone deacetylases 2 in steroid resistant childhood nephrotic syndrome

[Speaker] Harshit Singh:1
[Co-author] Narayan Prasad:2, Saurabh Chaturvedi:1, Vikas Agarwal:1, Kritika Singh:1
1:Clinical Immunology, Sanjay Gandhi Post Graduate Institute of medical Sciences, India, 2:Nephrology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, India

Background
Glucocorticoids are one of the most common and reliable therapy for inflammatory, autoimmune and proliferative diseases · Moreover¸ some patients show a poor or absent response · The activated glucocorticoid receptors interact with co-repressor molecules to impair NFΚB ¯associated coactivator activity ¸ which reduces histone acetylation · Reduction in histone acetylation occurs through the recruitment of histone deacetylase HDAC2 to the activated inflammatory gene complex by activated glucocorticoid receptor · To evaluate HDAC2 expression via immunohistochemistry and assess the HDAC2 mRNA expression in SR INS ·
Methods
Immunohistochemical HDAC2 expression was analyzed on renal biopsy samples in SR patients n=15 ¸M=9 ¸mean age 8.4±2.59 and healthy control HC n=6 ¸ M=3 ¸ mean age 20.47± 7.26 · Healthy control samples were taken from Nephrectomy specimen ¸ which were biopsy proven · Patients in sustained remission SS n=25 ¸ M=5 ¸ mean age 9.85±7.08 for at least 6 months without steroid were recruited · All definitions are as per the criteria of International Study of Kidney Diseases in Childhood ·
Immunohistochemistry was performed with formalin fixed, paraffin embedded tissue section following heat induced antigen retrieval in Tris EDTA buffer pH 8.6 ·
Peripheral blood mononuclear cells were isolated from heparinized blood · Total RNA was isolated using trizol method. Real time quantitative PCR was performed using light cycler LC480 using SYBR green PCR technology with SYBR premix Relative gene expression levels were calculated and normalized to the corresponding levels of the housekeeping gene (GAPDH) ·
Results
Demographic significant difference were found in S.Albumin SSNS=2.87±98 ¸ SRNS=2.27±79 ¸ p=0.012 and proteinuria SSNS=13.18±3.09 ¸ SRNS=284±193.45 ¸ p=0.001·
HDAC2 nuclear expression in healthy control and steroid dependent samples was significantly higher as compared to the nuclear expression of HDAC2 of SR patients86.6 percent¸ p=0.002 · HDAC2 mRNA expression remained similar in PBMCs of SS patients p=0.730 as compared to that of HC whereas in SR patients was significantly decreased p=0.023 ·
Conclusion
Lower HDAC2 nuclear and gene expression in SR patients suggests use of inducers, which might lead to restoration of glucocorticoid response and better management of patients ·
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