Navneet K. Dhillon, Ph.D. Division of Pulmonary and Critical Care University of Kansas Medical Center
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1 Navneet K. Dhillon, Ph.D. Division of Pulmonary and Critical Care University of Kansas Medical Center
2 About 37 million people living with HIV/AIDS worldwide
3 HIV-Related Cardio-pulmonary complications Chronic obstructive pulmonary disease (COPD) Coronary artery disease Pulmonary Arterial Hypertension (PAH)
4 1997 Swiss HIV Cohort 0.5% Opravil et. al, AJRCCM 2006/2008 French HIV Registry 0.5% Humbert et. al, AJRCCM /Sitbon O, AIDS 2008, UCSF study 15% ( RVSP > 35mmHg), 6.7% (RVSP > 40mmHg) Hsue et al, AIDS 2012 Spain Cohort - 9.9% (RVSP>35mmHg), 2.8% 40-65mmHg) Quezada et al, AIDS 2015 German Cohort 6.1% Schwarze-Zander et.al, HIV Medicine ~ fold higher prevalence in HIV patients compared with general population.
5 Percentage of HIV-PH with IVDU Risk Factor Himelman et al. Am J Cardiol., 1989 : 50% (n=3/6) Speich et al. Chest, 1991 : 83% (n=5/6) Petitpretz et al. Circulation, 1994 : 60% (n=12/20) Opravil et al, Am J Respir Crit Care Med.,1997 : 84% (n=16/19) Aguilar & Farber, Am J Respir Crit Care Med., 2000 : 100% (n=6, prostacyclin therapy) Sitbon et al. AIDS, 2008 : 51% Quezada et al. AIDS, 2012: 59% (n=17/39) Araujo et al. World journal of cardiology,2014 : 78% (n=14/18)
6 HIV+IVDU (opioids+/-cocaine) Endothelial dysfunction Smooth muscle dysfunction Smooth muscle dysfunction Ham 56 George et al. Am J Respir Crit Care Med Jun 1;185(11):
7 SIV- infected macaques
8 H&E H&E Factor VIII/SMA Factor VIII/SMA Hyperactive proliferative endothelial cells within plexiform lesions
9 How combination of HIV-1 infection and IVDU exposure exacerbates pulmonary arteriopathy????
10 illicit drugs gp-120, Nef illicit drugs
11 Additive increase in the proliferation of pulmonary SMCs in the presence of HIV- Tat and cocaine Activation of proliferative PDGF receptor axis without increase in PDGF-BB ligand Ligand independent activation of PDGFR Dhillon et al, Am. J Resp. Cell & Mol. Biol Dalvi et al, Am J Respir Cell Mol Biol. 2015
12 Smooth Muscle Dysfunction Anti-proliferative Bone Morphogenetic Protein receptor ( BMPR) signaling in Cocaine & HIV-Tat mediated Smooth Muscle Hyperplasia Role of mirnas in the regulation of BMPR-2 expression Role of macrophage derived extracellular vesicles on HIV-1 and /cocaine associated smooth muscle dysfunction
13 Smooth Muscle Dysfunction Anti-proliferative Bone Morphogenetic Protein receptor ( BMPR) signaling in Cocaine & HIV-Tat mediated Smooth Muscle Hyperplasia Role of mirnas in the regulation of BMPR expression Role of macrophage derived extracellular vesicles on HIV-1 and /cocaine associated smooth muscle dysfunction
14 Bone Morphogenetic Protein Receptors Transforming growth factor β Receptors BMPs signaling pathway Humpath.com - Human pathology
15 Dalvi.Dhillon, Arterioscler Thromb Vasc Biol. 2013
16
17 Dalvi.Dhillon, Arterioscler Thromb Vasc Biol. 2013
18
19 Dalvi.Dhillon, Arterioscler Thromb Vasc Biol. 2013
20
21 Cocaine : IP, 14mg/kg body weight for 21 days
22
23 Full media Serum-starved
24 Full media Serum-starved
25 RNA Protein
26 Dalvi.Dhillon, Arterioscler Thromb Vasc Biol Increased BMPR mrna Decreased BMPR protein
27 Smooth Muscle Dysfunction Anti-proliferative Bone Morphogenetic Protein receptor ( BMPR) signaling in Cocaine & HIV-Tat mediated Smooth Muscle Hyperplasia Role of mirnas in the regulation of BMPR expression Role of macrophage derived extracellular vesicles on HIV-1 and /cocaine associated smooth muscle dysfunction
28 Chinnappan et al, JAHA, 2018, In press
29 Inhibition of BMPR2 targeting mirnas prevented the cocaine & Tat mediated decrease in BMPR2 protein levels BMPR2 mrna levels BMPR2 Protein levels
30 Overexpression of BMPR2 targeting mirnas enhanced the cocaine & Tat mediated decrease in BMPR2 protein levels BMPR2 mrna level BMPR2 Protein level
31 Loss or gain of BMPR2 targeting mirnas on smooth muscle proliferation Inhibition of mirnas using antagomir overexpression of mirnas using mimics
32 Increased expression of BMPR2 targeting mirnas in pulmonary arterial smooth muscle cells from HIVtransgenic rats exposed to cocaine
33 MiRNA 216a mediates inhibition of BMPR2 expression by directly binding to its 3 UTR Chinnappan et al, JAHA, 2018
34 Inhibition of BMPR2 translation by mir-216 without any effect on RNA In-vitro translation assay Firefly luciferase protein levels Reporter RNA levels
35
36 Smooth Muscle Dysfunction Anti-proliferative Bone Morphogenetic Protein receptor ( BMPR) signaling in Cocaine & HIV-Tat mediated Smooth Muscle Hyperplasia Role of mirnas in the regulation of BMPR expression Role of macrophage derived extracellular vesicles on HIV-1 and /cocaine associated smooth muscle dysfunction
37
38 Increased perivascular inflammation around remodeled vessels SIV+morphine macaque HV Tg rat +cocaine
39 Do HIV-infection and cocaine mediated alterations in the macrophage-derived extracellular vesicles (EVs) promote hyperproliferation of pulmonary arterial smooth muscle cells?
40 Human monocyte derived macrophages Un-treated HIV-1 Cocaine (1µM) Collected supernatant day 2 or 4 postinfection Isolated Extracellular vesicles : Differential ultracentrifugation Effect of EVs on pulmonary smooth muscle proliferation HIV-1 +cocaine
41 Nanosight Analyses of macrophage derived extracellular vesicles
42 TEM WB for Exosomal markers LAMP-1 HSP-60
43
44 Control phalloidin (red) +Exosomes EVs labeled with PKH67 (green)
45 PCNA β-actin Con Coc HIV HCoc
46 Small RNA sequencing of RNA cargo revealed : Differentially regulated (>1.5 fold) : mirna HIV+cocaine group : 17 (p<0.05) HIV group : 4 (p<0.05) Cocaine group: 5 (p<0.05)
47 DIANA mir path
48 PTEN/Akt/mTOR signaling
49
50 PTEN TSC-2 β-actin TSC-1 β-actin P-AKT AKT P-p70S6K p70s6k
51 Uptake of Tx-Red SiRNA Exosomes by SMCs
52 Sharma et al. FASEB J Apr 19
53 RVSP Con EVs H+C EVs Con EVs H+C EVs WT HIV+cocaine
54
55 Lab members Pranjali Dalvi Himanshu Sharma Mahendran Chinnappan Stuti Agarwal Aradhana Mohan Zachery Harter Julie Allen Leslie Spikes Joel Mermis Kun Wang Xui Bing Haihua Gu Collaborators Augustine Choi (Weill Cornell Medical College) Susan Morgello (Mount Sinai, NY) Amy O Brien Ladner Paul Cheney Lane Christenson Ossama Tawfik KIDDRC Bio-informatics Core Funding NIH R01DA031589, R01DA034542, R01HL129875
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Effect of Cocaine on Human Immunodeficiency Virus Mediated Pulmonary Endothelial and Smooth Muscle Dysfunction
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