Genetic and Hormonal Contributions to Gene Expression in Immune Cells

The origins of sex differences in human disease are elusive, in part because of difficulties in separating the effects of sex hormones and sex chromosomes. To isolate the effects of testosterone, estradiol, and sex chromosome complement (XX or XY) in humans, we implemented a four-state, cross-sectional study design of trans- and cisgender individuals, the former receiving exogenous testosterone or estradiol for at least a year as part of their gender-affirming medical care. This study design allowed us to compare four combinations of sex hormones and sex chromosomes in humans: XX individuals treated with exogenous testosterone (n=21), XY individuals treated with exogenous estradiol (n=13), untreated XX individuals (n=20), and untreated XY individuals (n=15). We chose to focus on the human immune system because it displays an impressively broad range of sex-differential biology. We collected blood samples from each individual and isolated PBMCs the same day. We performed 10X Genomics 3' single-cell RNA-sequencing (scRNA-seq) on PBMCs from the 69 individuals. We generated 150-bp paired end reads from a total of 358,426 cells (with an average of 48,007 reads/cell) and identified 18 distinct immune cell types. We found that testosterone, estradiol, and sex chromosome complement have unique, non-overlapping effects on PBMC cell abundances and gene expression. ]]>
We recruited 34 transgender individuals from the Gender Multispecialty Service clinic at Boston Children's Hospital. In order to participate in this study, transgender individuals must have received exogenous estradiol or testosterone for at least a year as part of their medical care. We recruited 35 cisgender individuals from the general population. Cisgender individuals were required to be between the ages of 14-30 years. Exclusion criteria for all individuals were pregnancy, infection or vaccination within the preceding two weeks, and inability to provide consent. Cisgender individuals were also excluded if they had any medical diagnoses or used any prescription medications. We obtained written informed consent from each individual. If the individual was a minor, we obtained written informed assent from the individual and written informed consent from a guardian. ]]>
This study is an IRB-approved (P00029637), longitudinal, observational study including transgender youth. We started recruiting from the Gender Multispecialty Service clinic at Boston Children's Hospital on July 15, 2019; the data deposited here are inclusive through September 1, 2024]]>