CROI 2019 Abstract eBook

Abstract eBook

Poster Abstracts

277 TRANSCRIPTOME ANALYSIS IN HPV+/HIV+ TISSUE REVEALS MARKERS OF HPV-DEPENDENT DYSPLASIA Eva Riveira-Muñoz 1 , Ivan Galvan-Fermenia 2 , Rafael de Cid 2 , Antoni Tarrats 3 , Marta Piñol 3 , Francesc García-Cuyás 3 , Jose A. Este 1 , Roger Badia 1 , Guillem Sirera 3 , Ester Ballana 1 1 IrsiCaixa Institute for AIDS Research, Badalona, Spain, 2 Institute for Health Science Research Germans Trias i Pujol, Badalona, Spain, 3 Fundació Lluita Contra la Sida, Badalona, Spain Background: HPV is accepted today as the necessary but not sufficient etiological agent for anal and cervical neoplasia and HIV appears to be a cofactor in the association between HPV and cervical neoplasia. The objective of our study is the identification of a distinct transcriptomic signature that may serve as predictive markers of dysplasia and/or malignancy in HIV+/+HPV patients. Methods: The study includes a unique cohort of 25 HIV-1 infected individuals co-infected with HPV with a clinical follow-up for more than 20 years. Tissue samples from individuals with signs of a high degree of anal dysplasia were chirurgical collected, together with samples from normal tissue from the same individual as control. After RNA extraction and quality control, RNA library was constructed (Illumina TruSeq RNA stranded) and sequencing was performed (Novaseq, 30M reads/sample). Data analysis was performed as implemented in the computational workflow for the detection of differentially expressed genes and pathways from RNA-seq data. Read alignment and count quantification was conducted using the Rsubread package and the statistical analysis was performed using the edgeR package. The differential expression analysis uses the quasi-likelihood functionality of edgeR. Results: Whole transcriptome sequencing was performed to examine differential gene expression profiles, and to perform gene annotation based on gene ontology pathway information. Analyses were successfully performed on all 25 paired-ends samples with overall read mapping ratio above 95%. Thirty genes showed significant changes between biopsies showing a high degree of dysplasia and apparently healthy control biopsies. Hierarchic clustering of data demonstrated a clear discrimination between healthy and dysplasic tissues, indicating a common pattern of gene expression changes between individuals. The identified differentially expressed genes include chemokines, potential restriction factors, a miRNA and genes associated to cell proliferation and cell transformation. After filtering the results based on functionality, we selected for further validation a group of 15 genes that fulfill the criteria for becoming a biomarker. Conclusion: Our analysis allowed the identification of at least 15 potential predictive markers of anal dysplasia in co-infected HIV/HPV individuals. Characterization of the selected genes may result in the development of new therapeutic approaches to treat HIV/HPV induced malignancies. 278 HPV CLEARANCE AND REINFECTION IN 2 YEARS AFTER RANDOMIZATION TO CRYOTHERAPY OR LEEP Christine J. McGrath 1 , Anthony Cagle 1 , Hugo De Vuyst 2 , Troy Querec 3 , Elizabeth Unger 3 , Sharon A. Greene 1 , Sameh Sakr 4 , Marleen Temmerman 5 , Nelly R. Mugo 6 , Michael H. Chung 1 1 University of Washington, Seattle, WA, USA, 2 International Agency for Research on Cancer, Lyon, France, 3 CDC, Atlanta, GA, USA, 4 Coptic Hope Center, Nairobi, Kenya, 5 Aga Khan University, Nairobi, Kenya, 6 University of Washington in Kenya, Nairobi, Kenya

276 EXPLORING THE MICROBIOTA FOR THE DIAGNOSIS OF ANAL PRECANCEROUS LESIONS IN MSM Sergio Serrano-Villar 1 , Alfonso Cabello 2 , María José Gosalbes 3 , Matilde Sanchez-Conde 1 , Begoña Monge 1 , Jorge Diaz 1 , Talía Sainz 4 , Patricia Roiz 1 , María J. Vivancos-Gallego 1 , Maria Jesus P. Elias 1 , Jose L. Casado 1 , Ana Moreno 1 , Santiago Moreno 1 , José A. Pérez-Molina 1 1 Hospital Ramón y Cajal, Madrid, Spain, 2 Fundacion Jimenez Diaz, Madrid, Spain, 3 FISABIO, Valencia, Spain, 4 La Paz University Hospital, Madrid, Spain Background: Anal cancer is a leading neoplasia in HIV-infected MSM. The current screening strategy is based on the detection of high-degree squamous intraepithelial lesions (HSIL), using anal cytology. While this approach shows good sensitivity, the specificity is poor. We aimed to identify in MSM a set of anal-associated bacterial biomarkers for the diagnosis of biopsy-proven HSIL (bHSIL). Methods: Cross-sectional prospective study performed in HIV+ and HIV- MSM referred to a high-resolution anoscopy clinic. The primary outcome was the presence of bHSIL at the inclusion or during the previous year. We analyzed fecal and mucosal microbiota to search for biomarkers predictive of the presence of bHSIL. We collected fecal samples in specific containers and obtained anal mucosa specimens with an anal cytobrush. The V3-V4 region of the 16S rRNA gene was sequenced using the Illumina platform. We selected the biomarkers based on their LDA scores, AUC-ROC in logistic regression models and concomitant predictive value in feces. Results: We included 118 HIV+ and 33 HIV- MSM: 47 had bHSIL during the previous year and 12 at the moment of the inclusion. Differences in alpha and bacterial diversity were significant between mucosa and feces, but non-significant in the comparison by presence of bHSIL or HIV status. Linear discriminant analysis (LDA) effect size (LEfSe) revealed 40 biomarkers in mucosa and 53 in stools. After exploring the predictive value of the 15 taxa with greater LDA scores, we selected four taxa in anal samples. Each 25% increase in the abundance of the Ruminococcaceae NK4A214 group and Alloprevotella genus were associated with a 17% (p=0.041) and 8% (p=0.016) increased risk of bHSIL, respectively. The absence of Prevotella melanonigenica and Ruminococcaceae UCG-014 were predictive of bHSIL (OR 6.1, P=0.018 and OR 3.2, P=0.026, respectively). From 35 (94%) of false positive cytologic results, the combination of these four biomarkers reclassified to true negative 33 (94%), significantly improving the predictive performance of anal cytology alone to AUC 0.805. Conclusion: We found anal-associated bacteria indicative of higher risk of precancerous anal lesions, which combination was highly specific. The microbiota could be exploited as a complementary diagnostic tool for anal cytology to overcome the low specificity and high rate of false positive results of the current screening strategy for anal cancer screening.

Poster Abstracts

Background: Women with HIV are at increased risk of high-risk human papillomavirus (hrHPV) infection. We compared hrHPV clearance and reinfection in HIV-infected women randomized to cryotherapy or loop

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