Research Perspective Volume 13, Issue 18 pp 21814—21837

Development of infrastructure for a systemic multidisciplinary approach to study aging in retired sled dogs

Analysis of retrotransposons in somatic cells of aging dogs. (A) Schematic depiction of a hypothetical connection between retrotransposon expansion and inflammaging. Activity of the reverse transcriptase (RT) encoded by LINE-1 (L1) retrotransposons results in insertional mutagenesis and cGAS-STING-mediated interferon type I (IFN) response and inflammation. (B) Illustration of our approach assessing the association between retrotransposon content and aging. Mosaicism in retrotransposon content among blood cells during aging will be assessed through computational bioinformatic analysis of data from whole genome sequencing of DNA isolated from the same dog at 2-year intervals. Determination of germline SINE and LINE-1 contents for individual dogs (shown as “genotype” for individual dogs, A-D) will be followed by quantitation of novel somatic copies of retrotransposons (new insertions indicated as “somatic expansion”). This will allow us to establish a Blood Retrobiome Expansion index (BREX) reflecting the number of somatic integrations in each DNA sample that can be tracked over time (e.g., ΔBREX between years 1 and 4 (Y1-Y4) in the figure) and compared with biological age assessment approaches based on DNA methylation and senescence marker analyses.

Figure 3. Analysis of retrotransposons in somatic cells of aging dogs. (A) Schematic depiction of a hypothetical connection between retrotransposon expansion and inflammaging. Activity of the reverse transcriptase (RT) encoded by LINE-1 (L1) retrotransposons results in insertional mutagenesis and cGAS-STING-mediated interferon type I (IFN) response and inflammation. (B) Illustration of our approach assessing the association between retrotransposon content and aging. Mosaicism in retrotransposon content among blood cells during aging will be assessed through computational bioinformatic analysis of data from whole genome sequencing of DNA isolated from the same dog at 2-year intervals. Determination of germline SINE and LINE-1 contents for individual dogs (shown as “genotype” for individual dogs, A-D) will be followed by quantitation of novel somatic copies of retrotransposons (new insertions indicated as “somatic expansion”). This will allow us to establish a Blood Retrobiome Expansion index (BREX) reflecting the number of somatic integrations in each DNA sample that can be tracked over time (e.g., ΔBREX between years 1 and 4 (Y1-Y4) in the figure) and compared with biological age assessment approaches based on DNA methylation and senescence marker analyses.