This document provides a biographical sketch for Phillip B. Hylemon including: 1) His education, positions held, and honors received including his work in bile acid metabolism and gut bacteria. 2) Contributions to science including discovering pathways of bile acid metabolism by gut bacteria and bile acid activation of cell signaling pathways. 3) Current research support including grants to study role of bile acids and gut bacteria in disease and regulation of hepatic sphingosine kinase by bile acids.

1. OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015)
BIOGRAPHICAL SKETCH
Provide the follow ing information for the Senior/key personnel and other significant contributors.
Follow this format for each person. DO NOT EXCEED FIVE PAGES.
NAME: Hylemon, Phillip B.
eRA COMMONS USER NAME (credential, e.g., agency login): phylemon
POSITION TITLE: Professor of Microbiology and Immunology and Medicine
EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing,
include postdoctoral training and residency training if applicable. Add/delete rows as necessary.)
INSTITUTION AND LOCATION
DEGREE
(if
applicable)
Completion
Date
MM/YYYY
FIELD OF STUDY
Barton College, Wilson, North Carolina B.S. 05/’67 Biology and
Mathematics
Virginia Polytechnic Institute and State University,
Blacksburg, Virginia Ph.D. 09/’71 Microbiology
Medical College of Virginia, Virginia Commonwealth
University, Richmond, Virginia Postdoctoral 08/’73 Microbial Physiology
and Biochemistry
A. Personal Statement
The major objective of the current research application is to investigate the role of bile acid activated cell
signaling pathways in cholestasis using both in vitro and in vivo model systems (bile duct ligation and mdr2-/-
).
In our previous research, we discovered that conjugated bile acids (CBAs) activated the sphingosine-1-
phosphate (S1P) receptor 2 (S1PR2) in primary hepatocytes and cholangiocytes as well as
cholangiocarcinoma cell lines (rodents and humans). Activation of the S1PR2 by either S1P or CBAs activates
the ERK1/2 and AKT (insulin signaling pathway) which led to proliferation of cholangiocytes and
cholangiocarcinoma cell lines. Inhibition of the S1PR2 by either chemical antagonists or knock down by
shRNA markedly inhibited cell proliferation (2-4). In our most recent studies, we discovered that activation of
the S1PR2 by CBAs significantly up regulated nuclear sphingosine kinase 2 (SphK2) with increased levels of
nuclear S1P (4). It has been previously reported (Dr. Spiegel’s laboratory) that S1P is a powerful inhibitor of
histone deacetylases 1 and 2 (HDAC 1/2) which increases histone acetylation and gene expression. We have
preliminary data presented in this application that S1PR2 and SphK2 are significantly upregulated in bile duct
ligated mice and in cholangiocarcinoma (2). In contrast, our preliminary results show that bile duct ligation of
the S1PR2-/-
mouse shows a marked decrease in cholangiocyte proliferation, inflammation and fibrosis
markers. Therefore, we hypothesize that the S1PR2 is a key G protein coupled receptor (GPCR) regulating cell
proliferation, inflammation and fibrosis in the liver. If our hypothesis is confirmed, it may allow the scientific
community to target this receptor for treatment of various cholestatic liver diseases and possibly
cholangiocarcinoma. We have assembled an outstanding research team to study this new area of liver cell
biology and cholestasis research including: Huiping Zhou, Ph.D. (expertise in GPCRs, cholangiocytes,
cholangiocarcinoma and animal models); Sarah Spiegel, Ph.D.(expertise in sphingosine-1-phosphate,
sphingosine kinases and cell biology); Kazuaki. Takabe M.D., Ph.D. (expertise in liver cell biology, animal
models, surgery and clinical hepatology); and Phillip Hylemon, Ph.D. (bile acid cell signaling, steroid
biochemistry and hepatocyte cell biology).

2. In summary, I have a long track record in research in the bile acid field (continuously funded by the NIH since
1975) and working with a large research group. In this regard, I was the PI or Co-PI and a Program Project
Grant funded in this area from 1986-2006 (PO1-DK038030). I am very excited about the current research
project as the basic science information has the potential to be rapidly translated into clinical applications.
1: Nagahashi M, Takabe K, Liu R, Peng K, Wang X, Wang Y, Hait NC, Wang X, Allegood JC, Yamada A,
Aoyagi T, Liang J, Pandak WM, Spiegel S, Hylemon PB, Zhou H. Conjugated Bile Acid Activated S1P
Receptor 2 Is a Key Regulator of Sphingosine Kinase 2 and Hepatic Gene Expression. Hepatology. 2014 Nov
1. doi:10.1002/hep.27592. [Epub ahead of print] PubMed PMID: 25363242.
2: Liu R, Zhao R, Zhou X, Liang X, Campbell DJ, Zhang X, Zhang L, Shi R, Wang G, Pandak WM, Sirica AE,
Hylemon PB, Zhou H. Conjugated bile acids promote cholangiocarcinoma cell invasive growth through
activation of sphingosine-1-phosphate receptor 2. Hepatology. 2014 Sep; 60(3):908-18. PubMed PMID:
PubMed Central PMCID:PMC4141906.
3: Studer E, Zhou X, Zhao R, Wang Y, Takabe K, Nagahashi M, Pandak WM, Dent P, Spiegel S, Shi R, Xu W,
Liu X, Bohdan P, Zhang L, Zhou H, Hylemon PB. Conjugated bile acids activate the sphingosine-1-phosphate
receptor 2 in primary rodent hepatocytes. Hepatology. 2012 Jan;55(1):267-76. PubMed PMID: 21932398;
PubMed Central PMCID: PMC3245352.
4. Dent, P, Fang Y, Gupta S, Studer E, Mitchell C, Spiegel S, Hylemon PB. Conjugated bile acids promote
ERK1/2 and AKT activation via a pertussis toxin-sensitive mechanism in murine and human hepatocytes.
Hepatology 2005, Dec;42(6):1291-9. PMID 16317705
B. Positions and Honors
1973-77 Assistant Professor of Microbiology/Immunology, Virginia Commonwealth University
1977-85 Associate Professor of Microbiology/Immunology, Virginia Commonwealth University
1985- Professor of Microbiology/Immunology, Biochemistry (joint appointment) and Medicine (joint
appointment), Virginia Commonwealth University
Honors:
Winner of the Adolf Windaus Prize, 1990 (given by the Falk Foundation, Freiburg, Germany, for outstanding
publications in the field of bile acid research). 2.) Winner of the Virginia’s Outstanding Scientist Award for 1991
(given by the Science Museum of Virginia). 3.) School of Basic Health Sciences Outstanding Scholar Award,
1992,1993 and 1994. 4.) Winner of Virginia Commonwealth University’s Award of Excellence, 2005. This is the
highest honor given by VCU.
Other Experience and Professional Memberships:
American Academy of Microbiology, 1986-; President, Virginia Branch, American Society for Microbiology,
1986-88; American Chemical Society, 1992-2000; American Liver Foundation Grant Review Committee, 1991-
1993; 2015- ; Special Reviewer National Institutes of Health (Gastroenterology and Nutrition Division), 1986-
88; NIH Reviewer’s Pool-Metabolism Study Section, 1994-98; Special Reviewer for Xenobiotic and Nutrient
Disposition and Action and Hepatobiliary Pathophysiology (ZRG1) Study Section, 2009-present. NIH-NCI
Reviewer ad hoc, 2012-14. Editorial Board, Journal of Lipid Research 2005-present; Associate Editor, Journal
of Nutrition and Metabolism, 2010-present. Ad hoc reviewer for numerous scientific journals.
C. Contributions to Science
During my career, my laboratory has focused on two main areas of scientific research: 1) metabolism of
bile acids and steroid hormones by human gut bacteria and role of the human gut microbiome in
health and disease. In the late 1980’s, my laboratory discovered a new 8 step biochemical pathway (bile acid
7α/7β-dehydroxylation) found in specific human gut bacteria. This pathway produces secondary bile acids

3. (deoxycholic acid and lithocholic acid) from primary bile acids (cholic acid and chenodeoxycholic acid),
respectively. We subsequently discovered, cloned and characterized a large >11kb bile acid inducible (bai)
operon in Clostridium scindens encoding 8 genes. We then individually cloned expressed and characterized
each gene product (bile acid transporter and various enzymes in this pathway) (reviewed in a). We also
discovered that specific bile acid 7α/7β-dehydroxylating gut bacteria can convert glucocorticords to androgens
(b). Recently, in collaboration with a group at Scripps Institute, we obtained the 3D structure of bile acid 7α-
dehydratase, the rate limiting enzyme in this pathway (submitted). Finally, in collaboration with J.S. Bajaj,
M.D., we have reported evidence that bile acids help determine the structure of the human gut microbiome
(reviewed in c) and that secondary bile acids increases colonic inflammation in alcoholic cirrhosis patients (d).
Recently, we show that transfer of fecal samples from these patients to normal Germ-free mice mimics many
of the pathophysiological features found in these patients (submitted).
a. Ridlon JM, Kang DJ, Hylemon PB. Bile salt biotransformations by human intestinal bacteria. J. Lipid
Res. 2006 Feb;47(2)241-59 PMID 16299351
b. Ridlon JM, Ikegawa S, Alves JM, Zhou B, Kobayashi A, Iida, T, Mitamura K, Tanabe G, Serrano M, De
Guzman A, Cooper P, Buck GA, Hylemon PB. Clostridium scindens: a human gut microbe with a high
potential to convert glucocorticoids to androgens. J. Lipid Res. 2013 Sep;54(9)2437-49. PMC3735941
c. Ridlon JM, Alves JM, Hylemon PB, Bajaj JS. Cirrhosis, bile acids and gut microbiota: unraveling a
complex relationship. Gut Microbes 2013 Sep-Oct;4(5)382-7. PMC3839982
d. Kakiyama G, Hylemon PB, Zhou H, Pandak WM, Heuman DM, Kang DJ, Takei H, Nittono H, Ridlon
JM, Fuchs M, Gurley EC, Wang Y, Liu R, Sanyal AJ, Gillevet PM, Bajaj JS. Colonic inflammation and
secondary bile acids in alcoholic cirrhosis. Am J Physiol Gastrointest Liver Physiol 2014 Jun
1;306(11)G929-37. PMC4152166
2) I have also been working on the regulation of bile acid synthesis and bile acid activating cell
signaling pathways since the early 1980’s. We were one of the first laboratories to use primary rodent
hepatocytes to study the regulation of bile acid synthesis and were the first to show that cholesterol 7α-
hydroxylase (CYP7A1) was transcriptionally regulated by bile acids in vitro (a) and in vivo in the chronic bile
fistula rat. Our group was also the first laboratory to report that bile acids induced an “intestinal factor” that was
involved in the regulation of CYP7A1 using the chronic bile fistula rat model. We were the first laboratory to
report that bile acids activate the ERK1/2 pathway via the epidermal growth factor receptor (b). We also
showed that activation of the JNK1/2 signaling pathway down-regulated CYP7A1 (c) and that the bile acid
activated AKT pathway is linked to the activation of the nuclear receptor FXR (d). However, I think that our
recent discoveries of the activation of the S1PR2 by conjugated bile acids and that this receptor regulates the
levels and activity of nuclear SphK2 and hepatic gene expression may turn out to be the most important
contribution. We now hypothesize that this receptor is key to regulation of cell proliferation, inflammation and
fibrosis in the liver.
a. Hylemon PB, Gurley EC, Stravitz RT, Litz JS, Pandak WM, Chiang JY, Vlahcevic ZR. Hormonal
regulation of cholesterol 7 alpha-hydroxylase mRNA levels and transcriptional activity in primary rat
hepatocyte cultures. J Biol Chem 1992 Aug 25;267(24)16886-71 PMID1512229.
b. Rao YP, Studer EJ, Stravitz RT, Gupta S. Qiao L, Dent P, Hylemon PB. Activation of the Raf-
1/MEK/ERK cascade by bile acids occurs via the epidermal growth factor receptor in primary rat
hepatocytes. Hepatology 2002. Feb;35(2)307-14. PMID 11826403
c. Gupta S, Natarajan R, Payne SG, Studer EJ, Spiegel S, Dent P, Hylemon PB. Deoxycholic acid
activates the c-Jun N-terminal kinase pathway via FAS receptor activation in primary hepatocytes. Role
of acidic sphingomyelinae-mediated ceramide generation in FAS receptor activation. J. Biol Chem 2004
Feb 13;279(7)5821-8. PMID 14660582.
d. Cao R, Cronk ZX, Zha W, Sun L, Wang X, Fang Y, Studer E, Zhou H, Pandak WM, Dent P, Gil G,
Hylemon PB. Bile acids regulate hepatic gluconeogenic genes and farnesoid X recptor via G(alpha)i-
protein-coupled recptors and the AKT pathway. J. Lipid Res 2010 Aug;51(8)2234-44. PMC2903791

4. Complete List of Published Work in My Bibliography
http://www.ncbi.nlm.nih.gov/sites/myncbi/1Zqvb8-u-
qt5f/bibliography/47508026/public/?sort=date&direction=ascending
D. Research Support
Ongoing Research Support
1BX0013828 Hylemon (P.I.) 7/01/12-6/30/16
VETERANS ADMINISTRATION MERIT GRANT “ROLE OF BILE ACIDS AND GUT BACTERIA IN GI
DISEASES”
The overall aim of this grant is to determine the role of secondary bile acids and androgens generated from
C21-glucocorticoids by human gut bacteria play in the etiology of colon cancer and cholesterol gallstone
disease. Determine the role of gut bacteria in the pathophysiology of liver cirrhosis. Role P.I.
2R01 DK088664 Hylemon (P.I.) Zhou (P.I.) 5/01/12-04/31/16
NIH/NIDDK “REGULATION OF HEPATIC SPHK2 BY BILE ACIDS: EFFECTS ON LIPID METABOLISM. The
overall aim of this grant is to explore the role of bile acids in activation of sphingosine-1-phosphate signaling in
the liver and effects on lipid metabolism. Role Contact P.I.
1R01 DK104893 Zhou (P.I.) Hylemon (P.I.) 12/01/15-11/30/19
NIH/NIDDK “BILE ACID AND SPHINGOSINE-1-PHOSPHATE RECEPTOR-MEDIATED SIGNALING IN
CHOLESTASIS”. The overall aim of this grant is to investigate the role of conjugated bile acid activated
sphingosine-1-phosphate receptor 2 in liver injury and cholestasis. Role Co-PI.
Completed Research Support in last Five Years
RO1DK057543 Hylemon (PI) 5/01/07-03/30/12
NIH/NIDDK “BILEACID ACTIVATED CELL SIGNALING IN THE LIVER AND NUTRITION”. The overall aim of
this grant was to determine the role of bile acid cell signaling in the regulation of nutrient metabolism in the
liver.
R01AI057189 Hylemon (PI) 3/01/04-2/28/10
NIH/ NIAID “HIV Protease Inhibitors and Hepatic Lipid Dysregulation”. The overall aim of this grant was to
determine the mechanisms of lipid dysregulation by different HIV protease inhibitors in the liver.