The long-term objective of this proposal is to investigate the role of zinc finger transcriptional factor, Kruppel like factor-2 (KLF2) in the regulation of osteoclastogenesis and rheumatoid arthritis (RA)-related pathogenesis. Among the cellular components monocytes play major roles in mediating progression of RA. The monocytes migrate from the peripheral blood to the arthritic joint tissues and secrete pro-inflammatory factors, and these factors in turn mediate inflammation and recruit other immune cells, which mediate differentiation of monocytes towards osteoclasts. Since it has been reported that KLF2 can induce quiescence to the immune cells and also inactivate monocytes in response to stimulation (active monocytes are required to promote osteoclastogenesis) and because our preliminary results indicate severe arthritic joint changes in KLF2 hemizygous mice (KLF2 homozygous knock out mice are embryonically lethal), we therefore hypothesize that this transcription factor may inhibit osteoclastogenesis and thereby, pathogenesis of RA.
Aim1 will determine the importance of myeloid KLF2 in regulating the pathogenesis in RA. Experimental Design: Conditional gain and loss of gene function approach will be undertaken to determine the role of KLF2 in the regulation of pathogenesis in vivo. Complementary histologic, histomorphometric, immunohistochemical, and radiologic (micro CT) studies will be used to evaluate extent of the disease. Human monocytes and joint tissues from RA and healthy controls will be studied to confirm findings.
Aim 2 will examine the bases for KLF2's ability to regulate osteoclastogenesis. Experimental Design: Gain- and loss-of-function approaches will be used to determine the role of KLF2 in osteoclastic differentiation. Additionally, the cross talk between KLF2 and osteoclastogenesis markers (NFATc1, NF?B, Cathepsin K and MMP9) and osteoblastogenesis markers (RANK/RANKL, OPG, BMP2/4, and Runx2) will be determined. Human monocytes from RA and healthy controls will also be studied to confirm findings.
Aim 3 will investigate whether the effects of pharmacological compound on arthritis and osteoclastogenesis are KLF2 dependent. We found that a group of pharmacological compound, such as HDAC inhibitor (HDACi) induces KLF2 expression in myeloid cells. Experimental Design: We will verify if HDACi induce KLF2 in myeloid cells in mice. Next, we will determine if the HDACi-mediated regulation of inflammation and osteoclastogenesis is KLF2- dependent using various molecular approaches (structure-functions studies, co-immunoprecipitation, and ChIP assays). The knowledge generated from this study will not only identify a novel endogenous regulator of osteoclastogenesis in RA, but also indicate newer and effective strategies to control this disease.

Public Health Relevance

The macrophage is one of the critical regulators of the body's response to inflammation that contributes to the pathogenesis in arthritis. Recent studies from our group have identified a transcription factor KLF2 that regulates key aspects of macrophage biology. This proposal seeks to understand the mechanisms underlying the function of the transcription factor KLF2 with the goal of developing future alternate therapeutic approach by modulating KLF2 and their downstream targets for the treatment of rheumatoid arthritis in human.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
1R01AR068279-01A1
Application #
9096497
Study Section
Atherosclerosis and Inflammation of the Cardiovascular System Study Section (AICS)
Program Officer
Mao, Su-Yau
Project Start
2016-04-01
Project End
2016-08-31
Budget Start
2016-04-01
Budget End
2016-08-31
Support Year
1
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Ohio State University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Das, Manjusri; Laha, Dipranjan; Kanji, Suman et al. (2018) Induction of Kr├╝ppel-like factor 2 reduces K/BxN serum-induced arthritis. J Cell Mol Med :
Rolph, Daniela N; Deb, Moonmoon; Kanji, Suman et al. (2018) Ferutinin directs dental pulp-derived stem cells towards the osteogenic lineage by epigenetically regulating canonical Wnt signaling. Biochim Biophys Acta Mol Basis Dis :
Kanji, Suman; Das, Hiranmoy (2017) Advances of Stem Cell Therapeutics in Cutaneous Wound Healing and Regeneration. Mediators Inflamm 2017:5217967
Jha, Prerana; Das, Hiranmoy (2017) KLF2 in Regulation of NF-?B-Mediated Immune Cell Function and Inflammation. Int J Mol Sci 18: