{
  "abstract": "Introduction Data on HIV-1 reservoir dynamics of HIV during treatment with long-acting cabotegravir and rilpivirine (LA CAB/RPV) remain limited. This study aimed to investigate intra-patient viral evolution and the persistence of transcriptionally active proviruses, providing insights into reservoir stability and the impact of long-term LA CAB/RPV therapy.Methods Six people living with HIV initiating LA CAB/RPV at our centre were enrolled. Samples were collected at three different time points: before the first injection (T1), at the fourth injection (T4) and at the seventh injection (T7). The numbers of HIV-1-infected cells and the transcriptional activity were assessing by measuring cell-associated HIV-1 DNA (CAD) and cell-associated HIV-1 RNA (CAR) levels using digital PCR (dPCR) targeting three genomic regions (DNA: LTR-GAG-TREX1 and RT-GAG-TREX1; RNA: GAG-TAT-POLYA). Whole genome sequencing was performed at all time points. Two additional subjects experiencing virological failure on LA CAB/RPV were included as a comparator group.Results All participants were males, Italian born, with a median age of 47 years (interquartile range, IQR: 39–50 years) and a median duration of antiretroviral therapy of 13.95 years (IQR: 12–14 years). All subjects were MSM; 5 carried B and 1 CRF02_AG subtypes. At T7, integrase resistance-associated mutations were detected in one subject. About failing subjects, resistance-associated mutations were observed in one case while no resistance mutations were detected in the second subject at viral rebound. Off-target mutation analysis in gag and env genes revealed several variations, with the highest mutation burden in subjects with virological failure; only one gag mutation was consistently detected across all individuals.dPCR analyses showed a progressive decrease in quantification of single positive targets, both in DNA and RNA overtime. In contrast, for double and triple positive targets, the decrease was less pronounced, except for the DNA triplex with gag-trex1-rt targets. In the RNA triplex, with the tat-gag-poly-A targets, the levels at T1 and T4 were comparable, while at T7 a more marked reduction was observed. Subjects with virological failure showed higher CAD and CAR levels, although differences were not statistically significant. Triplex analyses were largely comparable between the two groups.Conclusion During the first year of treatment, LA CAB/RPV regimen significantly impacts the HIV-1 reservoir. CAD and CAR measurements showed concordant trends, with most participants showing zero long transcripts and intact proviruses. Whole genome analysis revealed previously undetected DNA mutations suggesting a certain dynamism in viral mutations even under effective long-acting treatment.",
  "authors": [
    {
      "affiliations": [
        "Department of Infectious Diseases, ASST Fatebenefratelli Sacco University Hospital, Milan, Italy"
      ],
      "name": "A Bergna"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "C della Ventura"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "I Serravalli"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "M Liturri"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "C Mauro"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "E Marinosci"
    },
    {
      "affiliations": [
        "Department of Infectious Diseases, ASST Fatebenefratelli Sacco University Hospital, Milan, Italy"
      ],
      "name": "MV Cossu"
    },
    {
      "affiliations": [
        "Department of Infectious Diseases, ASST Fatebenefratelli Sacco University Hospital, Milan, Italy"
      ],
      "name": "C Gervasoni"
    },
    {
      "affiliations": [
        "Department of Infectious Diseases, ASST Fatebenefratelli Sacco University Hospital, Milan, Italy",
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "S Antinori"
    },
    {
      "affiliations": [
        "Department of Infectious Diseases, ASST Fatebenefratelli Sacco University Hospital, Milan, Italy",
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "A Riva"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "G Zehender"
    },
    {
      "affiliations": [
        "Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy"
      ],
      "name": "A Lai"
    }
  ],
  "title": "OC16 Characterisation of HIV-1 reservoir in subjects on long-acting therapy",
  "uid": "fb1f9794-a528-515a-bb42-60175c6b18dc"
}