Alterations of adipose tissue induced by HIV or SIV infection: Tat and Nef-induced senescence and fibrosis

Jennifer GORWOOD1, Tina EJLALMANESH1, Matthieu MANTECON1, Michael ATLAN2, Jean-Philippe BASTARD1,3, Bruno FEVE1, Christine BOURGEOIS4,

Olivier LAMBOTTE4, Jacqueline CAPEAU1, Véronique BEREZIAT1 & Claire LAGATHU1

1Sorbonne Universités, UPMC Paris 06, Inserm UMR_S 938, ICAN, Paris, France2 APHP, Tenon Hospital, Department of Plastic surgery, Paris, France3 APHP, Tenon Hospital, Department of Biochemistry, Paris, France, 4Université Paris Sud, Inserm UMR_S 1184, Le Kremlin Bicêtre, France

Increased prevalence of comorbidities classically

associated with aging, such as cardio-metabolic diseases.

Trunk lipohypertrophy(SAT et VAT)

Aging and comorbidities in HIV-infected patients receiving ART

ART

HIV ?

FibrosisInflammation

Normal aging is also associated with adipose tissue redistribution, inflammation and fibrosis

Premature aging of the adipose tissue?

Adipocyte DysfunctionAdipogenesis

alterations

Caron-debarle M et.al, TMM, 2010Bastard JP et.al, Lancet, 2002

Adipose tissue is a reservoir for HIV/SIV

The adipose tissue is an HIV reservoir Damouche et al, PLOS Path, 2015Couturier et.al, AIDS, 2015

Adipose tissue is a reservoir for HIV/SIVBystander effect of HIV proteins

The adipose tissue is an HIV reservoir Damouche et al, PLOS Path, 2015Couturier et.al, AIDS, 2015

Hypothesis:

Hosted HIV/SIV could release viral proteins exerting a bystander effect on non-infected adipose stem cells, inducing inflammation, oxidative stress, senescence

and extracellular matrix (ECM) remodeling

Materiel & Methods 1: in vitro model, human ASCs isolated from abdominal SAT

Materiel & Methods 1: in vitro model of human ASCs isolated from abdominal SAT

Characteristics of a senescent cell

Results 1: Tat and Nef induce senescence in ASCs

Proliferation Senescence marker:SA-β-galactosidase activity

In response to HIV protein treatment:• Decreased proliferation• Increased number of senescent cells

• Increased expression of p21 and p16 and activation of P53• Increased IL-6 and IL-8 secretion

Results 2: Tat and Nef induce senescence in ASCsand senescence associated dysfunction

Cycle arrest proteins SASP : Senescence Associated Secretory Phenotype

Oxidative stress

Mitochondrial mass Mitochondrial membrane potential

Results 2: Tat and Nef induce senescence in ASCsand senescence associated dysfunction

• Tat and Nef induce oxidative stress and mitochondrial dysfunctions• Antioxidant treatment partially prevents Tat- and Nef- Induced senescence

Materiel & Methods 2: evaluation of adipogenesis

Results 3: Nef-induced senescence is associated with decreased adipogenesis

Lipid accumulation Adiponectin and leptin secretion

• Decreased lipid accumulation • Decreased expression of adipogenic markers and adipokine secretion• Altered expression of collagens I and VI

Adipogenic markers

Conclusion 1

Tat and Nef induce premature senescence of ASCOnly Nef affected adipogenesis

Different mechanisms of action ?

Materiel & Methods 1: In vivo model of Cymologus Macaques SAT and VAT

Cynomolgus macaque

(Macaca fascicularis)

In collaboration with CEA Primatology Centre and Pr R. Le Grand (CEA, France)

Adipose tissue

Non infected

Control(n=4)

Infected

SIV +(n=4)

Age 8,0± 0,9years

4,8± 3,8years

weight 10,3± 4,2kg

5,9± 1,6kg

Meanduration of

infection- 398 days

(SIVmac251)

Damouche et al, PLOS Path, 2015

Results 4: preliminary results SIV infection induce fibrosis and small adipocytes in SAT and VAT

in SAT and VAT from SIV + macaques• decreased adipocytes size

Damouche et al, PLOS Path, 2015

Results 4: preliminary results SIV infection induce fibrosis and small adipocytes in SAT and VAT

in SAT and VAT from SIV + macaques• decreased adipocytes size• Increased fibrosis and collagen VI expression

Conclusion 2

• Tat and Nef induce senescence in ASCs and alter adipogenesis

Conclusion 2

• Tat and Nef induce senescence in ASCs and alter adipogenesis• SIV/HIV participates in fibrosis and decrease adipocyte size in SAT & VAT

Conclusion 2

Adipose tissue dysfunction could contribute to cardio-metabolic disorders observed in HIV-infected patients

• Tat and Nef induce senescence in ASCs and alter adipogenesis• SIV/HIV participates in fibrosis and decrease adipocyte size in SAT & VAT

Primatecentre

CEA ImmunologyVirology

UMR1184CEA

Acquired and genetic

lipodystropiesUMRS 938St Antoine

centre

Pr Roger Le GrandNathalie Bosquet

Virgile MonetLeslie Gosse

Dr Christine BourgeoisPr Olivier Lambotte

Dr Véronique Béréziat Dr Claire Lagathu

Pr Jacqueline Capeau Prof Bruno Fève

Dr Jean-Philippe Bastard

Acknowledgments

Plastic Sugery

Tenon Hospital

Dr Michael Atlan

Implication of oxidative stress in Tat and Nef induced senescence

• NAC restores ASC proliferative capacity• NAC reduce the number of senescent cells

NAC : N-acétylcystéine, antioxydant

PDL SA-β-galactosidase activity

Nef- and Tat-induced senescence alters extracellular matrix remodelling

• During adipogenesis, Tat and Nef increase the expression of collagens I and VI and modify the fibronectinnetwork