Erin E. Sundermann, Ph.D. | 220 Dickinson Street Suite B | San Diego, CA, 92103 | P 858-552-8585 z7286 | [email protected]

Inflammation-Related Genes are Associated with Epigenetic Aging in HIV Erin E. Sundermann1, Mariam A. Hussain1,2, David J. Moore1, Steven Horvath3, David T.S. Lin4, Michael S. Kobor5, Andrew Levine3 1Department of Psychiatry, University of California, San Diego; 2San Diego State University/University of California San Diego Joint Doctoral Program in Clinical Psychology; 3Department of Neurology, 4 4 University of California, Los Angeles; , School of Public Health, University of California Los Angeles; The Centre for Molecular Medicine and Therapeutics, BC Children's Hospital HNRC • CHARTER • TMARC • CNTN Research Institute.

Background Statistical Analyses Results Results HIV & AGING • ANCOVAs were conducted to compare mean age acceleration z-scores • Genotypes associated with higher inflammatory levels in the IL-6 Figure 2. Percent with any HANA condition by IL-6 genotype • There is some evidence of “accelerated aging” in HIV+ persons between genotype group separately for each SNP. (minor allele carriers) and IL-10 genes (major allele homozygotes) group ○Age-related may be over-represented & appear 5-10 • Logistic regressions were conducted to compare odds of any past/current were associated with higher accelerated aging z-scores (ps<.05) p=.03 yrs earlier in HIV+ persons vs. general population (Pathai et al., HANA condition between genotype group separately for each SNP. • Any past/current HANA condition was over 3 times more likely in IL-6 80 2014). 70 • Covariates considered included demographics, non age-associated minor allele carriers vs. major allele homozygotes (OR=3.4, 60 • Chronic levels of inflammation and immune activation are amplified in comorbidities (e.g. substance use disorders, major depressive disorder, 95%CI=0.27-1.65, p=.03). 50 both HIV infection and aging. Thus, compounding effects of aging and Hepatitis C virus), HIV characteristics (e.g., duration of disease, • TNFα SNP was not significantly associated with accelerated aging or 40 HIV on inflammation may underlie accelerated aging in HIV+ persons. pre-mortem plasma CD4 & viral load levels), study site and assay batch. HANA. 30 20 GENETICS & AGING a % HANA with condition Figure 1. Mean accelerated aging z-scores by genotype group. 10 • DNA methylation (DNAm) levels increase with age and can be used as Table 1: Sample Characteristics 0 p=.005, Cohen’s d=0.28 an epigenetic biomarker of aging known as the 'epigenetic clock’ Major Allele Minor Allele Carriers Variables IL-6 IL-6 IL-10 IL-10 TNFα TNFα 0.4 (Horvath, 2013). Homozygous (n=32) Major Minor Major Minor Major Minor 0.2 (n=123) Allele Allele Allele Allele Allele Allele IL-6 genotype Score

Aim - Homo- Carriers Homo- Carriers Homo- Carriers 0 We examined whether three inflammation-related single nucleotide zygotes (n=32) zygotes (n=76) zygotes (n=28) -0.2 polymorphisms (SNPs) are risk factors for accelerated aging (as (n=123) (n=79) (n=125) determined by epigenetic clock) and HIV-associated non-AIDS Age at death 48.0 (9.6) 46.8 (8.6) 47.8 (8.7) 47.8 (10.1) 48.2 (8.9) 46.6 (11.5) Accelerated Z Aging -0.4 Conclusions (HANA) conditions among HIV+ persons. Major Allele Minor Allele Carriers Male sex, % 76.4% 84.4% 83.5% 72.3% 75.2% 89.3% Homozygous (n=32) • Alterations in the production/degradation of IL-6 and IL-10 (n=123) cytokines may contribute to accelerated aging in HIV. Methods Education 12.8 (2.7) 12.0 (2.9) 12.9 (2.7) 12.3 (2.7) 12.7 (2.6) 12.3 (3.0) IL-6 genotype • Genetic polymorphisms in the interleukin pathway may be PARTICIPANTS Self-reported white, used to identify HIV+ persons at high risk for accelerated 155 post-mortem HIV+ cases from the National NeuroAIDS Tissue 73.2 71.9 81.0b 64.4b 71.2c 78.6c p=.02, Cohen’s d=0.35 aging and to inform therapeutic strategies for improving health • (Hisp. & non-Hisp.)% 0.4 Consortium (NNTC; Request #s: R334, R432, R432Ra) span in HIV+ persons. Lifetime diagnosis of 40.4 37.0 37.5 41.9 42.0 25.0 Score 0.2

• Year of death ranged from 1999 to 2014 - • Our sample lacked HIV-uninfected controls. Thus, it remains MDD, % 0 to be determined whether the IL-6 and IL-10 risk alleles have INFLAMMATION-RELATED SNPs b b c c Lifetime diagnosis of 47.0 53.8 56.1 40.0 53.5 28.0 additive or synergistic effects with HIV infection on • Interluekin pathway: IL-6 rs1800796 (-174G>C) & IL-10 rs1800872 (- substance -0.2 Accelerated Accelerated Z Aging inflammaging. 592C>A) abuse/dependence, % -0.4 • As the HIV+ population ages, future research should examine • Tumor Necrosis Factor alpha (TNFα) rs1800629 (-308G>A) Pre-mortem HAND, % 77.2 75.0 79.7 73.7 75.2 82.1 Major Allele Minor Allele Carriers whether findings generalize to older HIV+ persons. • SNP genotype dichotomized into major allele homozygotes vs. minor Homozygous (n=76) Hepatitis C virus, % 36.5 40.9 35.4 39.6 35.4 39.6 (n=79) allele carriers IL-10 genotype ACCELERATED AGING MEASURE (Epigenetic clock) Nadir CD4 count 59.8 41.6 59.6 (71.3) 51.9 (68.9) 59.3 (74.2) 42.7 (46.1) (cells/μl) (71.2) (64.3) References • DNAm levels of 353 CpGs measured in occipital lobe brain tissue with 0.4 p≥.05, Cohen’s d=0.35 • Horvath S & Levine AJ. J Infect Dis; 2015; 212(10):1563-73. Illumina Infinium HumanMethylation450 BeadChip Pre-mortem CD4 131.7 104.8 101.7 151.0 120.7 155.8 count (cells/μl) (178.5) (134.9) (138.8) (195.2) (151.0) (241.1) Score - • Pathai S et al.. Genome Biol 2013; 14:R115. • Accelerated aging defined by the difference between observed DNAm 0.2 3.8 (1.5) 3.7 (1.6) 4.0 (1.6) 3.6 (1.5) 3.5 (1.6) 3.8 (1.5) levels (predicted age) and that predicted by a linear model in age- Pre-mortem log plasma viral load 0 matched controls Est. duration of HIV 12.2 (6.7) 10.7 (5.2) 12.3 (5.6) 11.5 (7.2) 11.9 (6.6) 12.0 (5.7) Accelerated Z Aging -0.2 + z-score value = accelerated aging compared to controls disease (years) -0.4 HANA CONDITIONS Pre-mortem ART 82.9 84.4 82.3 85.5 84.0 78.6 Major Allele Minor Allele Carriers Presence of past or current cerebrovascular disease, liver disease, status, % prescribed • Homozygous (n=28) kidney disease, COPD, , and diabetes was either self-reported in- Note. Values represent M (SD) unless otherwise note aSignificant difference between IL-6 b c (n=125) life or extrapolated from medical records. genotype groups. Significant difference between IL-10 genotype groups. Significant TNFα genotype difference between TNFα genotype groups. MDD = Major Depressive Disorder. HAND = HIV- associated neurocognitive disorder. ART = antiretroviral therapy. HNRP.HIVRESEARCH.UCSD.EDU