Role of myeloid KLF2 in the neonatal innate immune response

This proposal describes a five-year mentored plan for research training and career development that will facilitate the development of Dr. Devashis Mukherjee as an independent investigator in the pathogenesis of immunological effects of sepsis in preterm neonates. This plan will build on Dr. Mukherjee’s prior research training as a transcription factor biologist and his clinical experience as a neonatologist to investigate the role of myeloid-KLF2 in the pathogenesis of the dysregulated innate immune response responsible for the high mortality burden of preterm neonatal sepsis. Case Western Reserve University and Rainbow Babies and Children’s Hospital rank among the top pediatric institutions in the United States in terms of NIH funding, research output, and patient care excellence. In addition, Dr. Mukherjee will train under the mentorship of the Dr. Alex Huang and Dr. George Dubyak, who are renowned immunologists with a highly successful track record of mentorship. Gram-negative sepsis in a preterm neonate is highly fatal, with 1 in 2 of these babies dying despite timely initiation of antimicrobial treatment. Dr. Mukherjee has identified a critical transcription factor, Kruppel-like factor (KLF) -2, which decreases in the myeloid cells during gram-negative sepsis and elevates the immune system into pro- inflammatory overdrive through increased NLRP3 signaling and increased preponderance of aged, pro- inflammatory neutrophils in the circulation. Dr. Mukherjee will investigate the mechanistic basis of these findings through a combination of murine genetic models and human translational studies with two specific aims: Aim 1. Investigate how decreased KLF2 affects the neonatal neutrophil response to gram negative sepsis by increased NLRP3 signaling. Aim 2: Determine how decreased KLF2 at an earlier postnatal age leads to persistence of aged circulating neutrophils via cortical actin reorganization. These mechanistic studies will be critical in understanding the role of KLF2 in the dysregulated preterm immune response leading to the overwhelmingly high mortality in this vulnerable population. In addition, Dr. Mukherjee will gain significant training under the mentorship of Dr. Huang and Dr. Dubyak in neutrophil and inflammasome biology, advanced imaging techniques such as two-photon microscopy, advanced flow cytometry, and neutrophil functional assays. He will also receive training on murine models of bacterial sepsis under Dr. Jennifer Bermick at the University of Iowa, on chromatin immunoprecipitation and bioinformatic analysis under Dr. Berkley Gryder at CWRU, and on reporter assays to investigate transcription factor binding to promoter sites under Dr. Mukesh Jain at Brown University and Dr. Lalitha Nayak at Indiana University. This will lay the foundation for KLF2-targeted therapies as possible adjunctive treatments for sepsis in preterm neonates and Dr. Mukherjee’s transition to an independent investigator in developmental and myeloid cell biology. Project Number: 1K08AI198077-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Allergy and Infectious Diseases (NIAID) | Principal Investigator: Devashis Mukherjee | Institution: CASE WESTERN RESERVE UNIVERSITY, CLEVELAND, OH | Award Amount: $165,240 | Activity Code: K08 | Study Section: Special Emphasis Panel[ZRG1 IVBH-A (91)] View on NIH RePORTER: https://reporter.nih.gov/project-details/1K08AI19807701A1