title: Genomic structure and expression of the human serotonin 2A receptor gene (HTR2A) locus: identification of novel HTR2A and antisense (HTR2A-AS1) exons.
abstract: BACKGROUND
  The serotonin 2A receptor is widely implicated in genetic association studies and remains an important drug target for psychiatric, neurological, and cardiovascular conditions. RNA sequencing redefined the architecture of the serotonin 2A receptor gene (HTR2A), revealing novel mRNA transcript isoforms utilizing unannotated untranslated regions of the gene. Expression of these untranslated regions is modulated by common single nucleotide polymorphisms (SNPs), namely rs6311. Previous studies did not fully capture the complexity of the sense- and antisense-encoded transcripts with respect to novel exons in the HTR2A gene locus. Here, we comprehensively catalogued exons and RNA isoforms for both HTR2A and HTR2A-AS1 using RNA-Seq from human prefrontal cortex and multiple mouse tissues. We subsequently tested associations between expression of newfound gene features and common SNPs in humans.
  
  
  RESULTS
  We find that the human HTR2A gene spans ~66 kilobases and consists of 7, rather than 4 exons. Furthermore, the revised human HTR2A-AS1 gene spans ~474 kilobases and consists of 18, rather than 3 exons. Three HTR2A exons directly overlap with HTR2A-AS1 exons, suggesting potential for complementary nucleotide interactions. The repertoire of possible mouse Htr2a splice isoforms is remarkably similar to humans and we also find evidence for overlapping sense-antisense transcripts in the same relative positions as the human transcripts. rs6311 and SNPs in high linkage disequilibrium are associated with HTR2A-AS1 expression, in addition to previously described associations with expression of the extended 5' untranslated region of HTR2A.
  
  
  CONCLUSIONS
  Our proposed HTR2A and HTR2A-AS1 gene structures dramatically differ from current annotations, now including overlapping exons on the sense and anti-sense strands. We also find orthologous transcript isoforms expressed in mice, providing opportunities to elucidate the biological roles of the human isoforms using a model system. Associations between rs6311 and expression of HTR2A and HTR2A-AS1 suggest this polymorphism is capable of modulating the expression of the sense or antisense transcripts. Still unclear is whether these SNPs act directly on the expression of the sense or antisense transcripts and whether overlapping exons are capable of interacting through complimentary base-pairing. Additional studies are necessary to determine the extent and nature of interactions between the SNPs and the transcripts prior to interpreting these findings in the context of phenotypes associated with HTR2A.
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. ruble_cara_lee_ann@lilly.com.
authors.forename: Cara L
authors.initials: CL
authors.lastname: Ruble
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. smith_ryan_matthew@lilly.com.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Smith
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. calley_john_n@lilly.com.
authors.forename: John
authors.initials: J
authors.lastname: Calley
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. munsie_leanne_m@lilly.com.
authors.forename: Leanne
authors.initials: L
authors.lastname: Munsie
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. airey_david_charles@lilly.com.
authors.forename: David C
authors.initials: DC
authors.lastname: Airey
authors.affiliation: Lieber Institute for Brain Development, Baltimore, MD, ᅟ. garygao@gmail.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Lieber Institute for Brain Development, Baltimore, MD, ᅟ. JooHeon.Shin@libd.org.
authors.forename: Joo Heon
authors.initials: JH
authors.lastname: Shin
authors.affiliation: Lieber Institute for Brain Development, Baltimore, MD, ᅟ. thomas.hyde@libd.org.|Departments of Neurology, Psychiatry and Behavioral Sciences, John Hopkins University School of Medicine, Baltimore, MD, ᅟ. thomas.hyde@libd.org.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Hyde
authors.affiliation: Lieber Institute for Brain Development, Baltimore, MD, ᅟ. richard.straub@libd.org.
authors.forename: Richard E
authors.initials: RE
authors.lastname: Straub
authors.affiliation: Lieber Institute for Brain Development, Baltimore, MD, ᅟ. drweinberger@libd.org.|Departments of Psychiatry, Neurology, Neuroscience, and the Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, ᅟ. drweinberger@libd.org.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Weinberger
authors.affiliation: Tailored Therapeutics, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, IN, ᅟ. nisenbaum_laura_k@lilly.com.
authors.forename: Laura K
authors.initials: LK
authors.lastname: Nisenbaum
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D016373:DNA, Antisense; D005091:Exons; D006801:Humans; D051379:Mice; D020641:Polymorphism, Single Nucleotide; D017397:Prefrontal Cortex / Q000378:metabolism; D022821:RNA Splice Sites; D044402:Receptor, Serotonin, 5-HT2A / Q000235:genetics*; D012559:Schizophrenia / Q000235:genetics; D016415:Sequence Alignment; D014158:Transcription, Genetic
chemical_list: D016373:DNA, Antisense; D022821:RNA Splice Sites; D044402:Receptor, Serotonin, 5-HT2A