Department of Medicine

University of Pittsburgh

Division of Pulmonary, Allergy, and Critical Care Medicine
3459 Fifth Avenue,
628 NW
Pittsburgh, PA 15213
Academic Office: (412) 692-2210
Comprehensive Lung Center (Patient Care and Referral): (412) 648-6161

Rama K. Mallampalli, MD

Professor and Vice Chair for Research
Department of Medicine

Director, Acute Lung Injury Center of Excellence
of the University of Pittsburgh

UPMC Montefiore Hospital - NW628
3459 Fifth Avenue
Pittsburgh, PA 15213

Phone: 412-624-8735
Fax: 412-383-7113
Email: mallampallirk@upmc.edu
Administrative Assistant: Mary Pat Rocco
Administrative Assistant Email: roccomp@upmc.edu
VAPHS Assistant: Pamela Constable
VAPHS Assistant Email: Pamela.Constable@va.gov

Bio

Dr. Mallampalli received his MD from the University of Wisconsin. He completed an internship and residency in Internal Medicine at Hennepin County Medical Center in Minneapolis, MN where he also served as chief medical resident. Dr. Mallampalli completed a Pulmonary and Critical Care fellowship at the University of Iowa, where he obtained his research training in surfactant lipids. At Iowa, Dr. Mallampalli was appointed Professor of Medicine and Biochemistry and served as Associate Chair. Dr. Mallampalli has had or currently holds editorial appointments with several journals including the Journal of Biological Chemistry, the American Journal of Physiology, and the American Journal of Respiratory and Critical Care Medicine. He also serves on multiple peer review panels including the LIRR Study Section and Program Project Grant applications for the NIH. He is an elected member of the Association of American Physicians and the American Society of Clinical Investigation, an Established Investigator of the American Heart Association, and a Career Investigator of the American Lung Association. In September of 2009, Dr. Mallampalli was recruited as Chief of the Pulmonary Division of the VA Pittsburgh Healthcare System and as the Director of the Acute Lung Injury Center at the University of Pittsburgh. He currently serves as Vice Chair for research in the Department of Medicine.

Academic and Research Interests

Dr. Mallampalli’s research in the area of pulmonary molecular and cell biology as it relates to acute lung injury (ALI) and the mechanisms of sepsis. He is an internationally recognized investigator in the area of lipid metabolism and ubiquitin-mediated proteolysis as it relates to inflammation and injury.

The primary goal of his research is to better understand inflammation, that when left unchecked, leads to tissue and organ injury. His laboratory discovered a critical role for ubiquitin E3 ligase components in the molecular control of inflammation and how they impact regulatory proteins in sepsis and pneumonia. His current activity investigates the discovery, characterization, and biological role of orphan ubiquitin E3 ligase subunits belonging to the Skp-Cullin1-F box (SCF) family that control site-specific ubiquitination and disposal of key target proteins involved in innate immunity, inflammation, and cellular lifespan. His laboratory recently discovered a new model of innate immunity that led to the synthesis of the first genus of ubiquitin E3 ligase (F box) small molecule inhibitors that modulate proteolysis thereby inhibiting inflammation in preclinical models of sepsis and multi-organ failure.

Figure 1. Molecular regulation of inflammation through pro-inflammatory cytokines mediated by F box proteins. Microbial infection or stimuli can robustly activate a variety of cell surface receptors linked to TRAF proteins that serve as critical intermediary signaling proteins to mediate cytokine synthesis and release. The F box protein Fbxl2 serves as a sentinel inhibitor of TRAFs by mediating their polyubiquitination (red circles) and proteasomal degradation in cells. Fbxl2 specifically targets at a conserved tryptophan domain within all TRAF 1-6. During microbial infection, another F box protein, Fbxo3, targets Fbxl2 for its ubiquitination and degradation at K201; this process is facilitated by glycogen synthase kinase (GSK3b) phosphorylation (green circle) of Fbxl2 at T404. Wild-type Fbxo3 in this pathway potently activates cytokine driven inflammation, whereas a naturally occurring Fbxo3V221I polymorphism is hypofunctional. A novel small molecule Fbxo3 inhibitor, BC-1215, reduces inflammation by antagonizing actions of Fbxo3 on TRAF–cytokine signaling.

Dr. Mallampalli’s second area of research interest is in the investigation of the mitochondrial-specific phospholipid, cardiolipin. His laboratory uncovered a new paradigm for pneumonia. Pneumonia patients had increased levels of a toxin, cardiolipin, which reproduces this disorder when given to mice. From clues in a rare disease, his group discovered that a pump normally removes cardiolipin from lung fluid but is degraded in pneumonia. These observations may be transformative in that they could lead to non-antibiotic therapies in this illness.

Figure 2. Cardiolipin, elevated in pneumonia causes lung injury.
Quantification of cardiolipin in subjects with pneumonia. A. Median (gray line) and distribution (black circles) of cardiolipin abundance in tracheal aspirates from subjects with nonpulmonary critical illness (NPCI, n = 5), pneumonia (PNA, n = 17) and CHF (n = 6). B. MicroCT scan images were obtained on live mice (in vivo) 1 h after i.t. administration of cardiolipin (50 nmol, (low), 100 nmol (high)) versus control mice (top images).

A third focus is to investigate the molecular mechanisms for control of major phospholipids of animal membranes and of lung surfactant, including phosphatidylcholine (PC). PC levels are tightly controlled, in part, by the rate-regulatory phosphoenzyme cytidylyltransferase (CCT). His work previously investigated the molecular physiology of how CCT is controlled by reversible phosphorylation events within its carboxyl-terminus and its regulation by enzyme turnover. In models of inflammatory lung injury, surfactant PC biosynthesis is impaired because CCT activity decreases as a result of post-translational enzyme modification and gene transcriptional repression. Specifically, he discovered that CCT is coordinately degraded by calpains and the ubiquitin system in models of pulmonary sepsis. However, CCT is also protected during lung inflammation by the stabilizing ligand, calmodulin.

Figure 3. Calmodulin (CaM) Binds CCTα In Vivo. A. Mammalian 2-hybrid assay. Cells were co-transfected with CCTα-Gal4BD (CCTα) and CaM-Gal4AD (CaM) plasmids as fusion proteins separately [inset] or in combination with a plasmid construct encoding a b-galactosidase reporter gene (pG5CAT). Cells were lysed and assayed for b-galactosidase activities. B. FRET Analysis. Cells were transfected with YFP-CaM and CFP-CCTα and CaM-CCTα interaction at the single cell level was imaged using laser scanning microscopy before and after photobleaching. Shown in the upper sets of panels is single cell imaging showing that after acceptor photobleaching, fluorescence intensity of YFP decreased and CFP increased, confirming protein interaction between CaM and CCTα. Below: the same FRET was confirmed quantitatively by graphing of fluorescence intensities.

Key Publications

Chen BB, Coon TA, Glasser JR, McVerry BM, Zhao J, Zhao Y, Zou C, Ellis B, Sciurba FC, Zhang Y, and Mallampalli RK A combinatorial F box protein directed pathway controls TRAF stability to regulate inflammation. Nat. Immunol. 2013 March 31. PMID: 23542741

Ray NB, Durairaj L, Chen BB, McVerry BJ, Ryan AJ, Donahoe M, Waltenbaugh AK, O'Donnell CP, Henderson FC, Etscheidt C, McCoy D, Agassandian M, Hayes-Rowan E, Coon TA, Butler TL, Gakhar L, Mathur SN, Sieren JC, Tyurina YY, Kagan VE, McLennan G, and Mallampalli RK. Dynamic regulation of cardiolipin by the lipid pump, ATP8b1, determines the severity of lung injury in experimental pneumonia. Nat. Med. 16:1120-27, 2010.

Chen BB, Glasser JR, Coon TA, and Mallampalli RK. F box protein FBXL2 exerts human lung tumor suppressor-like activity by ubiquitin-mediated degradation of cyclin D3 resulting in cell cycle arrest. Oncogene, 31: 2566-2579, 2012.

Agassandian M, Chen BB, Pulijala R, and Mallampalli RK. Calcium-calmodulin kinase I cooperatively regulates nucleocytoplasmic shuttling of CCTα by accessing a nuclear export signal. Mol Biol Cell. 2012 Jul; 23(14):2755-69. Epub 2012 May 23. PMID: 22621903

Chen BB, Glasser JR, Coon TA, Zou C, Miller HL, Fenton, M, McDyer JF, Boyiadzis M, and Mallampalli RK. F box protein FBXL2 targets cyclin D2 for ubiquitination and degradation to inhibit leukemic cell proliferation. Blood 119 (13): 3132-3141, 2012, 2012.

Zhao J, Wei J, Mialki RK, Chen BB, Coon C, Zou C, *Mallampalli RK, Zhao Y. F-box protein FBXL19-mediated ubiquitination and degradation of the IL-33 receptor limits pulmonary inflammation. Nat. Immunol. 2012 Jun 3;13(7):651-8. *co-senior author.

Chen BB, Coon TA, Glasser JR, and Mallampalli RK. Calmodulin antagonizes a calcium-activated SCF ubiquitin E3 ligase subunit, FBXL2, to regulate surfactant homeostasis. Mol. Cell. Biol. 31: 1905-1920, 2011.

PubMed Link

News

Read highlights of Dr. Mallampalli's work in Nature Reviews Immunology and Nature Reviews Drug Discovery