Tomo Tarui1, Aimee Kim2, Alan Flake2, Lauren McClain2, John D Stratigis2, Inbar Fried3, Rebecca Newman4, Donna K Slonim4, Diana W Bianchi5. 1. Mother Infant Research Institute, Pediatrics, Floating Hospital for Children, Tufts Medical Center, Boston, MA. Electronic address: ttarui@tuftsmedicalcenter.org. 2. Center for Fetal Research, Pediatric Surgery, Children's Hospital of Philadelphia, Philadelphia, PA. 3. Department of Biomedical Informatics, Harvard Medical School, Boston, MA. 4. Department of Computer Science, Tufts University, Boston, MA. 5. Medical Genetics Branch, National Human Genome Research Institute, Bethesda, MD.
Abstract
BACKGROUND: Cell-free RNA in amniotic fluid supernatant reflects developmental changes in gene expression in the living fetus, which includes genes that are specific to the central nervous system. Although it has been previously shown that central nervous system-specific transcripts are present in amniotic fluid supernatant, it is not known whether changes in the amniotic fluid supernatant transcriptome reflect the specific pathophysiologic condition of fetal central nervous system disorders. In myelomeningocele, there is open communication between the central nervous system and amniotic fluid. OBJECTIVES: The purpose of this study was to identify molecular pathophysiologic changes and novel disease mechanisms that are specific to myelomeningocele by the analysis of amniotic fluid supernatant cell-free RNA in fetuses with open myelomeningocele. STUDY DESIGN: Amniotic fluid supernatant was collected from 10 pregnant women at the time of the open myelomeningocele repair in the second trimester (24.5±1.0 weeks); 10 archived amniotic fluid supernatant from sex and gestational age-matched euploid fetuses without myelomeningocele were used as controls (20.9±0.9 weeks). Differentially regulated gene expression patterns were analyzed with the use of human genome expression arrays. RESULTS: Fetuses with myelomeningocele had 284 differentially regulated genes (176 up- and 108 down-regulated) in amniotic fluid supernatant. Known genes that were associated with myelomeningocele (PRICKLE2, GLI3, RAB23, HES1, FOLR1) and novel dysregulated genes were identified in association with neurodevelopment and neuronal regeneration (up-regulated, GAP43 and ZEB1) or axonal growth and guidance (down-regulated, ACAP1). Pathway analysis demonstrated a significant contribution of inflammation to disease and a broad influence of Wnt signaling pathways (Wnt1, Wnt5A, ITPR1). CONCLUSION: Transcriptomic analyses of living fetuses with myelomeningocele with the use of amniotic fluid supernatant cell-free RNA demonstrated differential regulation of specific genes and molecular pathways relevant to this central nervous system disorder, which resulted in a new understanding of pathophysiologic changes. The data also suggested the importance of pathways that involve secondary disease, such as inflammation, in myelomeningocele. These newly identified pathways may lead to hypotheses that can test novel therapeutic targets as adjuncts to fetal surgical repair.
BACKGROUND: Cell-free RNA in amniotic fluid supernatant reflects developmental changes in gene expression in the living fetus, which includes genes that are specific to the central nervous system. Although it has been previously shown that central nervous system-specific transcripts are present in amniotic fluid supernatant, it is not known whether changes in the amniotic fluid supernatant transcriptome reflect the specific pathophysiologic condition of fetal central nervous system disorders. In myelomeningocele, there is open communication between the central nervous system and amniotic fluid. OBJECTIVES: The purpose of this study was to identify molecular pathophysiologic changes and novel disease mechanisms that are specific to myelomeningocele by the analysis of amniotic fluid supernatant cell-free RNA in fetuses with open myelomeningocele. STUDY DESIGN: Amniotic fluid supernatant was collected from 10 pregnant women at the time of the open myelomeningocele repair in the second trimester (24.5±1.0 weeks); 10 archived amniotic fluid supernatant from sex and gestational age-matched euploid fetuses without myelomeningocele were used as controls (20.9±0.9 weeks). Differentially regulated gene expression patterns were analyzed with the use of human genome expression arrays. RESULTS: Fetuses with myelomeningocele had 284 differentially regulated genes (176 up- and 108 down-regulated) in amniotic fluid supernatant. Known genes that were associated with myelomeningocele (PRICKLE2, GLI3, RAB23, HES1, FOLR1) and novel dysregulated genes were identified in association with neurodevelopment and neuronal regeneration (up-regulated, GAP43 and ZEB1) or axonal growth and guidance (down-regulated, ACAP1). Pathway analysis demonstrated a significant contribution of inflammation to disease and a broad influence of Wnt signaling pathways (Wnt1, Wnt5A, ITPR1). CONCLUSION: Transcriptomic analyses of living fetuses with myelomeningocele with the use of amniotic fluid supernatant cell-free RNA demonstrated differential regulation of specific genes and molecular pathways relevant to this central nervous system disorder, which resulted in a new understanding of pathophysiologic changes. The data also suggested the importance of pathways that involve secondary disease, such as inflammation, in myelomeningocele. These newly identified pathways may lead to hypotheses that can test novel therapeutic targets as adjuncts to fetal surgical repair.
Authors: Asli Sirmaci; Seyra Erbek; Justin Price; Mingqian Huang; Duygu Duman; F Başak Cengiz; Güney Bademci; Suna Tokgöz-Yilmaz; Burcu Hişmi; Hilal Ozdağ; Banu Oztürk; Sevsen Kulaksizoğlu; Erkan Yildirim; Haris Kokotas; Maria Grigoriadou; Michael B Petersen; Hashem Shahin; Moien Kanaan; Mary-Claire King; Zheng-Yi Chen; Susan H Blanton; Xue Z Liu; Stephan Zuchner; Nejat Akar; Mustafa Tekin Journal: Am J Hum Genet Date: 2010-05-06 Impact factor: 11.025
Authors: K Kato; T Kishi; T Kamachi; M Akisada; T Oka; R Midorikawa; K Takio; N Dohmae; P I Bird; J Sun; F Scott; Y Miyake; K Yamamoto; A Machida; T Tanaka; K Matsumoto; M Shibata; S Shiosaka Journal: J Biol Chem Date: 2001-02-05 Impact factor: 5.157
Authors: Enrico Danzer; Liping Zhang; Antoneta Radu; Michael W Bebbington; Kenneth W Liechty; N Scott Adzick; Alan W Flake Journal: Am J Obstet Gynecol Date: 2011-02 Impact factor: 8.661
Authors: N Scott Adzick; Elizabeth A Thom; Catherine Y Spong; John W Brock; Pamela K Burrows; Mark P Johnson; Lori J Howell; Jody A Farrell; Mary E Dabrowiak; Leslie N Sutton; Nalin Gupta; Noel B Tulipan; Mary E D'Alton; Diana L Farmer Journal: N Engl J Med Date: 2011-02-09 Impact factor: 91.245
Authors: David M Howard; Oliver Pain; Ryan Arathimos; Miruna C Barbu; Carmen Amador; Rosie M Walker; Bradley Jermy; Mark J Adams; Ian J Deary; David Porteous; Archie Campbell; Patrick F Sullivan; Kathryn L Evans; Louise Arseneault; Naomi R Wray; Michael Meaney; Andrew M McIntosh; Cathryn M Lewis Journal: Hum Mol Genet Date: 2022-02-21 Impact factor: 6.150