Basic Science Research Effort at Division of Cardiology
Laboratory Location: Medical Research Building (MRB 7.166)
Laboratory Telephone: 409.772.5870, 5871, 5872, and 5873.
Members of Cardiology Basic Science Research Laboratory
Ken Fujise, MD
Professor of Medicine and of Biochemistry and Molecular Biology
Nong Gao He
Research Associate III
Decha Pinkaew, Ph.D.
Fei Miao, Ph.D.
Runglawan Silakit, Ph.D.
Fortilin and Atherosclerosis:
Atherosclerosis is the leading cause of death in the US. Despite
extensive use of treatment regimens aimed at optimizing lipid levels,
about half of patients with acute myocardial infarction seen in
emergency departments have normal lipid profiles, suggesting that the
eradication of atherosclerosis and its complications calls for not only
statin therapy but also the development of innovative treatment
regimens. Since its inception in 1997, Fujise's lab has studied fortilin
and its role in vascular biology. The lab first reported that fortilin
is an anti-apoptotic molecule that interacts with MCL1, an
anti-apoptotic Bcl-2 family member protein critical for macrophage
survival. Furthermore, the lab showed that fortilin message levels are
significantly higher in patients with coronary artery disease (CAD); and
that atherosclerotic plaques over express fortilin. In addition, the
lab discovered that DHA, an artemisinin derivative, binds to and
facilitates degradation of fortilin. In the laboratory, we explore (a)
the basic mechanism of action of fortilin, (b) the role of fortilin in
atherosclerogenesis, (c) the use of fortilin as a biomarker of coronary
artery disease, (d) the use of DHA-derivatives for the prevention and
treatment of atherosclerosis. Research on fortilin and atherosclerosis
is currently supported
by NHLBI R01 (2013-2018), American Heart Association Grant-in-Aid
(2011-2013) and others.
Fortilin As a Biomarker of In Vivo Apoptosis
of cells undergo apoptosis each day in the average normal adult. The
ability to readily assess the degree of apoptosis in human diseases is
hampered by the lack of sensitive and specific serum biomarkers of
apoptosis. Fortilin is a novel prosurvival molecule that protects cells
against various noxious stimuli. While fortilin is secreted into the
extracellular space under certain conditions, the relationship between
the serum concentration of fortilin and the presence and extent of
apoptosis in vivo remains unknown. Using a newly developed fortilin
ELISA system, we have shown that fortilin exists in the normal human and
mouse circulation. In addition, fortilin serum levels are significantly
elevated in patients with solid cancer, in response to anti-cancer
chemo- or radiation therapy. The elevation of fortilin serum levels is
more robust and sensitive than that of such previously-reported serum
biomarkers of apoptosis as fragmented cytokeratin-18, cytochrome c, and
nucleosomal DNA. Further, targeted apoptotic liver damage induced by Jo2
anti-Fas (CD95) antibody consistently and significantly increases serum
fortilin levels in C57BL/6J mice. Finally, when challenged by
anti-human-Fas IgM antibody, Jurkat leukemic T cells apoptose and
release fortilin into the medium before plasma membrane integrity
becomes compromised. Conclusions: Taken together, our research suggest
that serum fortilin levels reflect the degree and extent of apoptosis
occurring in vivo. Fortilin is a viable serum biomarker of in vivo
apoptosis and can be utilized to noninvasively assess the status of in
vivo apoptosis in humans.
Fortilin and Cancer
Tumor suppressor protein p53 keeps us free of cancer when it is
function. Mutated p53 genes are seen in more than 50% of all human
cancers, making them the most frequently observed genetic derangement in
human cancer. At a molecular level, the ability of p53 to eliminate
cancerous cells relies on its ability to induce apoptosis, through
either the transcriptional activation of proapoptotic genes such as Noxa
, PUMA, and Bax or the
direct transcription-independent activation of Bax on mitochondria.
Growing cancers manage to keep p53 in check either by mutating the p53
gene itself or by expressing p53 inhibitors such as Mdm2. Recently, our
laboratory reported that fortilin is upregulated in many human
malignancies and that fortilin binds specifically to the
sequence-specific DNA binding domain of p53. The interaction of fortilin
with p53 blocks p53-induced transcriptional activation of Bax.
Fortilin, but not a double point mutant of fortilin lacking p53 binding,
inhibits p53-dependent apoptosis. Furthermore, cells with wild-type
p53 and fortilin, but not cells with wild-type p53 and the double-mutant
of fortilin lacking p53 binding, fail to induce Bax gene and apoptosis,
leading to the formation of large tumors in athymic mice. Fortilin is a
novel p53-interacting molecule and p53 inhibitor. Fortilin thus is a
logical molecular target in cancer therapy. Efforts to identify small
molecule inhibitors of fortilin is underway in our laboratory.
HPV and Heart Disease
Inflammation contributes to the development and progression of
atherosclerosis. In order to examine the association between human
papillomavirus (HPV) with cardiovascular disease (CVD), the data from
the National Health and Nutritional Examination Survey (2003-2006, 2450
women) was investigated. A total of 60 females (39 women were HPV DNA
positive, whereas 21 were negative) had coronary artery disease.
Presence of vaginal HPV DNA was associated with CVD. Odds ratio (OR) of
CVD comparing women with presence of vaginal HPV DNA to those without
was 2.30 (95% confidence interval [CI]: 1.27 to 4.16) after controlling
for demographics, health/sex behaviors, medical comorbidities,
cardiovascular risk factors, and management. At the same level of
adjustment, OR of CVD comparing women with cancer-associated HPV types
to those with negative HPV was 2.86 (95% CI: 1.43 to 5.70). HPV
infection, especially cancer-associated oncogenic types, is associated
with CVD among women. (J Am Coll Cardiol 2011;58:2001-6) Further studies
are necessary to establish the causal relationship between HPV
infection and atherosclerogenesis.
Garcinia dulcis, a plant that belongs to the Guttiferae family, is
widely distributed in Thailand, and other Southeast Asian countries. It
has been used in traditional medicine for centuries to treat various
inflammatory conditions. The main constituent of the plant is
morelloflavone, a biflavonoid comprising two covalently linked
flavones-apigenin and luteolin. The Fujise Lab tested if morelloflavone
prevented post-angioplasty restenosis and atherosclerosis, using mouse
models of post-angioplasty restenosis and atherosclerosis, respectively.
It was shown that morelloflavone blocks both restenosis and
atherosclerosis through the blockade of vascular smooth muscle cell
migration from the media to the intima.
- Methods And Compositions Relating To Fortilin, An Anti-Apoptotic Molecule, And Modulators Of Fortilin,USPO: 10/021,753 (Awarded)
- Composition and Method for Treatment and Prevention of Restenosis, USPO: 10/448,664 (Awarded)
- Reducing Myocardial reperfusion injury by the
combination therapy of protein kinase A activation and beta-1-adrenergic
receptor blockade (US Patent Pending, 12/997,485)
- Uses of morelloflavone (US Patent Pending, 12/931,310)
- Sinthujaroen P, Wanachottrakul N, Pinkaew P, Petersen J,
Phongdara A, Sheffield-Moore M, Fujise K. Elevation of serum fortilin
levels is specific for apoptosis and signifies cell death in vivo.
Biochimica et Biophysica Acta Clinical (BBA Clinical) 2014 in press
- Pinkaew D, Le RJ, Chen Y, Eltorky M, Teng BB, Fujise K.
Fortilin Reduces Apoptosis in Macrophages and Promotes Atherosclerosis.
American journal of Physiology Heart and Circulatory Physiology 2013.
- Chen Y, Fujita T, Zhang D, Doan H, Pinkaew D, Liu Z, Wu
J, Koide Y, Chiu A, Lin CJC, Chang JY, Ruan KH, Fujise K: The physical
and functional antagonism between tumor suppressor protein p53 and
fortilin, an anti-apoptotic protein. Journal of Biological Chemistry
2011. 286; 37: 32575-85. PMID: 21795694
- Kuo HK, Fujise K: Human papillomavirus and
cardiovascular disease among U.S. women in the National Health and
Nutrition Examination Survey, 2003-2006. Journal of American College of
Cardiology 2011: 58: 2001-6. PMID: 22032713
- Pinkaew D, Hutadilok-Towatana N, Teng BB, Mahabusarakam
W, Fujise K. Morelloflavone, a biflavonoid inhibitor of
migration-related kinases, ameliorates atherosclerosis in mice.
American Journal of Physiology-Heart and Circulation. 2011. In press
- Koide Y, Tonganunt M, Kato Y, and Fujise K.
Fortilin-deficiency causes embryonic lethality through the
overactivation of BMP4 Pathway. Biochimica et BiophysicaActa
2009;1790:326-38 PMID: 2695948
- Graidist P, Yazawa M, Tonganunt M, Nakatomi A, Lin CC,
Chang J, Phongdara A, Fujise K. Fortilin binds Ca2+ and blocks
Ca2+-dependent apoptosis in vivo. Biochemical Journal 2007;
408(2):181-91. PMID: 2267342
- Graidist P, Fujise K, Wanna W, Sritunyalucksana K,
Phongdara A: Establishing A Role For Shrimp Fortilin In Preventing Cell
Death. Aquaculture 2006; 255:157-164. ISSN: 0044-848
- Graidist P, Phongdara A, and Fujise K: Anti-apoptotic
protein partners fortilin and MCL1 independently protect cells from
5-FU-induced cytotoxicity: Fortilin-MCL1 interaction in apoptosis.
Journal of Biological Chemistry 2004; 279:40868-40875. PMID: 15262975
- Mnjoyan ZH, Dutta R, Zhang D, Teng BB, and Fujise K:
Paradoxical upregulation of tumor suppressor protein p53 in
serum-stimulated vascular smooth muscle cells. Circulation 2003;
108:4:464-471. PMID: 12860918
- Tulis DA, Mnjoyan ZH, Schiesser R, Shelat HS, Zoldehelyi
P, Fujise K: Adenoviral gene transfer of fortilin attenuates neointima
formation through suppression of vascular smooth muscle cell
proliferation and migration. Circulation 2003; 107:98-105. PMID:
- Zhang D, Li F, Mnjoyan ZH, Weidner D, Fujise K: Physical
and functional interaction between MCL1 and fortilin. The potential
role of MCL1 as a fortilin chaperone. Journal of Biological Chemistry
2002; 227 (40):37430-37438. PMID: 12149273
- Li F, Zhang D and Fujise K: Characterization of
fortilin, a novel anti-apoptotic protein. Journal of Biological
Chemistry 2001;276(50):47542-47549. PMID: 11598139
- Fujise K, Zhang D, Liu JL and Yeh ETH: Regulation of
apoptosis and cell cycle progression by MCL1. Differential role of MCL1.
Journal of Biological Chemistry 2000, 275(50):39458-65. PMID: 10978339
- Gong L, Kamitani T, Fujise K, Caskey LS, Yeh ETH:
Preferential interaction of sentrin with a ubiquitin-conjugating enzyme,
Ubc9. Journal of Biological Chemistry: 1997, 272(45)28198-201. PMID:
- Fujise K, Stacy L, Beck P, Yeh ETH, Chuang A, Brock TA,
Willerson JT: Differential effects of endothelin receptor activation on
cyclic flow variations in rat mesenteric arteries. Circulation 1997;
96(10);3641-6. PMID: 9396466
- Fujise K, Revelle M, Stacy L, Willerson JT, Beck P: A
tPA/P-selectin fusion protein is an effective thrombolytic agent.
Circulation 1997; 95:715-722. PMID: 9024162