Molecular Genetics of Growth and Development

Group Heads:

PostDocs:

• Dr. Kit Doudney
• Dr. David Monk
• Dr. Irwin Pauws
• Dr. Sayeda Abu-Amero

Research Assistants/PhD Students:

• Artemis Andreou
• Zoreh Motamedi

Principal Aims
To identify and study genes that are involved in key processes of fetal development that when disrupted cause common human disorders

The Role of Imprinted Genes In Fetal Growth and Human Reproduction.
Recently there has been growing interest in the concept of "genomic imprinting" and the knowledge that different genes or regions of chromosomes are active or inactive depending on their parental origin. It also seems clear from mouse studies that the early development of the trophoblast is paternally controlled and that of the embryo maternally controlled. Mouse studies creating disomic regions of known chromosome pairs have shown that the phenotype of mice exhibiting selected regions of uniparental disomy (UPD) was abnormal and often growth related.  As a consequence of the observations in mouse and UPD being found associated with several disease phenotypes, we are studying human intrauterine growth retardation (IUGR) for UPD and genomic imprinting effects. One model of particular interest to us is Silver-Russell syndrome in which the main feature is IUGR.  We have also extended analysis to preimplantation embryos specifically studying expression and imprinting of growth factor families. At present we are focusing on fetal growth with a particular interest in the role of the placenta.  To facilitate this we have recently collected a large cohort of mother-father-baby trios representing consecutive consenting births at Queen Charlotte’s and Chelsea Hospital.  The collection encompasses a normal distribution of low through to high birthweight babies and samples collected include blood and placenta to allow for both DNA and RNA analysis.

Key Publications
Lighten AD, Hardy K, Winston RML and Moore GE. IGF2 is parentally imprinted at the onset of expression during human preimplantation embryogenesis. Nat Genet (1997) 15: 122-123.

Preece MA, Price S, Davies V, Gough L, Stanier P, Trembath R and Moore GE. Maternal uniparental of chromosome 7 and the Silver Russell syndrome. J Med Genet (1997) 34: 6-9.

Monk D, Wakeling EL, Proud V Hitchins M, Abu-Amero SN, Stanier P, Preece MA and Moore, G.E. Duplication of 7p11.2-p13, including GRB10, in Silver-Russell syndrome. Am J Hum Genet (2000) 66: 36-46.

Arnaud P, Monk D, Hitchins M, Gordon E, Dean W,  Beechey CV, Peters J, Craigen W, Preece MA, Stanier P, Moore GE and Kelsey, G.  Conserved methylation imprints in the human and mouse GRB10 genes with divergent allelic expression suggests differential reading of the same mark. Hum Mol Genet (2003) 12:1005-1019.

Monk D, Smith R, Arnaud P, Preece MA, Stanier P, Beechey CV, Peters J, Kelsey G and Moore GE.  Imprinted methylation profiles for proximal mouse chromosomes 11 and 7 as revealed by methylation-sensitive representational difference analysis.  Mamm Genome (2003) 14: 805-816.

Hitchins M, and Moore GE. Genomic imprinting and fetal growth and development. Expert Rev Mol Med (2002) 1-19


Cleft palate
Clefts of the lip and/or palate (CL/P) are among the most common birth defects worldwide occurring with a prevalence of approximately 1 per 700 births. The majority are non-syndromic where CL/P occurs in isolation of other phenotypes. The etiology is complex, involving both major and minor genetic influences with variable interaction from environmental factors. Consequently, facial clefts usually occur either as isolated cases or with a family history but with no clear inheritance pattern.  However, by making use of DNA from a large Icelandic family with an unusual form of the disorder inherited as a single gene defect, we demonstrated that the mutated gene responsible for X-linked secondary cleft palate (CPX) mapped to Xq21.  This was one of the first localisations of a single gene causing a congenital malformation.  Extensive genetic, physical and transcriptional mapping techniques recently allowed us to identify the T-box transcription factor, TBX22 as the causative gene, finding a splice site mutation in the Icelandic family.  This gene was then investigated in additional CPX families, revealing further splice site, frame shift, nonsense and missense mutations.  All of the missense mutations occur in highly conserved residues of the T-box domain, the region required for binding to the DNA target sequences.  Our current work is to investigate the function of TBX22 during palate formation and identify downstream targets, which must also play a critical role in craniofacial development.

Key Publications
Braybrook C, Doudney K, Marçano ACB, Arnason A, Bjornsson A, Patton MA, Goodfellow PJ, Moore GE and Stanier P.  X-linked cleft palate and ankyloglossia (CPX) is caused by mutations in the T-box transcription factor gene TBX22. Nat Genet (2001) 29:179-183.

Braybrook C, Lisgo S, Doudney K, Henderson D, Marçano ACB, Strachan T, Patton MA, Villard L, Moore GE, Stanier P and Lindsay S. Craniofacial expression of human and murine TBX22 correlates with the cleft palate and ankyloglossia phenotype observed in CPX patients. Hum Mol Genet (2002) 11: 2793-2804.

Marçano ACB, Doudney K, Braybrook C, Squires R, Patton MA, Lees M, Richieri-Costa A, Lideral AC, Murray JC, Moore GE and Stanier P. TBX22 mutations are a frequent cause of cleft palate. J Med Genet (2004) 41: 68-74.

Stanier P and Moore GE. Genetic basis for cleft lip and palate: syndromic genes contribute to the incidence of nonsyndromic clefts. Hum Mol Genet (2004) 13: R73-R81.

Neural Tube Defects
Neural tube defects (NTD) as a group rank amongst the commonest congenital disorders in man (~1/1000 births). The underlying mechanisms causing NTD are difficult to study in the human because of the early stage of pregnancy at which they arise and because the mode of inheritance is complicated. However, studying simple animal models for NTD conditions might elucidate such mechanisms. The phenotype of the homozygous mouse mutants loop-tail (Lp), circletail (Crc) and Crash (Crsh) all affect the initial closure event in neural tube formation.  This results in a defect very similar to the severe human condition, cranio-rachischisis, where the neural tube (future spine) remains completely open and leads to fatality at or around the time of birth. In collaboration with the research team of Professor Andrew Copp at the Institute of Child Health (UCL, London), we successfully identified the mutant genes in each mouse.  Two of the genes encode homologues of the Drosophila planar cell polarity (PCP) proteins strabismus/vang (Vangl2) and flamingo/starry night (Celsr1). These transmembrane proteins are thought to interact in a multiprotein complex including frizzled and dishevelled, that determines cell polarity in the floor plate region, directing cell movements allowing for convergent extension and axial elongation in the developing embryo.  The Crc mutation was found in a gene called Scribble, also involved in cell polarity although not previously described in the PCP pathway.  We have shown that all three genes interact and any combination of double heterozygotes result in the severe NTD phenotype. Our current research is oriented towards investigating the biochemical pathways through which these genes function and investigating the potential role of PCP genes in human NTD.

Key Publications
Murdoch JN, Doudney K, Paternotte C, Copp AJ and Stanier P.  Severe neural tube defects in the loop-tail mouse result from mutation of Lpp1, a novel gene involved in floor plate specification.  Hum Mol Genet (2001) 10: 2593-2601.

Murdoch JN, Henderson D, Doudney K, Gaston C, Patternote C, Arkell R, Stanier P, and Copp AJ. Disruption of scribble (Scrb1) causes severe neural tube defects in the circletail mouse. Hum Mol Genet (2003) 12: 87-98.

Curtin JA, Quint E, Tsipouri V, Arkell RM, Cattanach B, Copp AJ, Henderson DJ, Spurr N, Stanier P, Fisher EM, Nolan PM, Steel KP, Brown SDM, Gray IC, Murdoch JN.  Mutation of Celsr1 disrupts planar polarity of inner ear hair cells and causes severe neural tube defects in the mouse.  Curr Biol (2003) 13: 1129-1133

Other Recent Publications
Moore GE, Abu-Amero SN, Wakeling EL, Bell G, Wilson A, Stanier P, Jauniaux E and Bennett ST.  Evidence that insulin is imprinted in the human yolk sac.  Diabetes (2001) 50: 193-203.

Mergenthaler S, Hitchins MP, Blagitko-Dorfs N, Monk D, Wollmann HA, Ranke MB, Ropers H-H, Apostolidou S, Stanier P, Preece MA, Kalscheuer VA, Thomas Eggermann and Moore GE, (The European Silver-Russell syndrome Consortium).  Conflicting reports of imprinting status of human GRB10 in developing brain. Am J Hum Genet (2001) 68: 543-544.

Carrel T, Herman GE, Moore GE and Stanier P.  Lack of mutations in ZIC3 in three X-linked pedigrees with neural tube defects. Am J Med Genet.(2001) 98: 283-285.

Hitchins MP, Monk D, Bell G, Ali Z, Wakeling EL, Preece P, Stanier P and Moore GE.  Human GRB10 is repressed on the maternal allele in the fetal central nervous system; evaluation of the role of GRB10 in Silver-Russell syndrome.  Eur J Hum Genet (2001) 9: 82-90.

Braybrook C, Warry G, Howell G, Arnason A, Bjornsson A, Ross MT, Moore GE and Stanier P.  Identification and characterisation of KLHL4, a novel human homologue of the Drosophila kelch gene that maps within the X-linked cleft palate and ankyloglossia (CPX) critical region. Genomics (2001) 72: 128-136.

Doudney K, Murdoch J, Patternote C, Bentley L, Gregory S, Copp AJ and Stanier P.  Comparative physical and transcript map of ~1 Mb around the loop-tail gene on distal mouse chromosome 1 and human chromosome 1q22-q23.  Genomics (2001) 72: 180-192.

Monk D, Hitchins M, Russo S, Preece M, Stanier P and Moore GE.  No evidence for mosaicism in Silver Russell syndrome.  J Med Genet. (2001) 38: e11.

Braybrook C, Warry G, Howell G, Mandryko V, Arnason A, Bjornsson A, Ross MT, Moore GE and Stanier P.  Physical and transcriptional mapping of the X-linked cleft palate and ankyloglossia (CPX) critical region. Hum Genet (2001) 108: 537-545.

Murdoch JN, Rachel RA, Shah S, Beerman F, Stanier P, Mason CA and Copp AJ.  Circletail, a new mouse mutant with severe neural tube defects: Chromosomal localisation and interaction with the loop-tail mutation. Genomics (2001) 78: 55-63.

Hitchins M, Stanier P, Preece M and Moore GE.  Silver-Russell syndrome: a dissection of the genetic aetiology and candidate chromosomal regions.  J Med Genet (2001) 38: 810-819.

Sharp A, Moore GE. and Eggermann T.  Evidence for skewed X inactivation for the involvement of trisomy 7 mosaicism in the aetiology of Silver-Russell syndrome. Eur J Hum Genet (2001) 9: 887-891.

Hitchins MP, Abu-Amero S, Apostolidou S, Monk D, Stanier P, Preece MA and Moore GE.  Investigation of the GRB2, GRB7 and CSH1 genes as candidates for the Silver-Russell syndrome (SRS) on chromosome 17q. J Med Genet (2002) 39: e13.

Eggermann T, Zerres K, Eggermann K, Moore GE and Wollmann HA. Clinical indications for uniparental disomy (UPD) testing in growth retarded patients Eur J Paediatr (2002) 161: 305-312.

Doudney K, Murdoch J, Braybrook C, Paternotte C, Bentley L, Copp AJ and Stanier P.  Cloning and characterisation of Igsf9 in mouse and human: a new member of the immunoglobulin superfamily expressed in the developing nervous system. Genomics (2002) 79: 663-670.

Nakabayashi K, Bentley L, Hitchins MP, Mitsuya K, Meguro M, Minagawa S, Bamforth JS, Stanier P, Preece MA, Weksberg R, Oshimura M, Moore GE and Scherer SW.  Identification of an imprinted antisense RNA (MESTIT1) in the human MEST locus on chromosome 7q32. Hum Mol Genet (2002) 11: 1743-1756.

Rogner CU, Danoy P, Matsuda F, Moore GE, Stanier P and Avner P.  Mutational analysis of NAP1L2 in neural tube defect patients reveals a cluster of SNPs within a CpG island that is highly conserved in the mouse orthologue.  Am J Med Genet (2002) 110: 208-214.

Monk D, Bentley L, Beechey C, Hithins M, Preece MA, Stanier P and Moore GE.  Characterisation of the growth regulating gene IMP3, a candidate for Silver-Russell syndrome.  J Med Genet (2002) 39: 575-581.

Monk D, Bentley L, Hitchins M, Clayton-Smith J, Ismail I, Stanier P, Preece MA and Moore GE.  Chromosomal abnormalities in the Silver Russell syndrome critical region on human chromosome 7p. Hum Genet (2002) 111: 376-387.

Hitchins MP, Monk D, Bentley L, Beechey C, Peters J, Kelsey G, Ishino F, Preece MA, Stanier P and Moore GE.  DDC and COBL are biallelically expressed genes flanking the imprinted gene GRB10 on 7p11.2-p12. Mammalian Genome (2002) 13: 686-691.

Murdoch JN, Doudney K, Gerrelli D, Wortham N, Paternotte C, Stanier P and Copp AJ.  Genomic organisation and embryonic expression of Igsf8, a novel immunoglobulin superfamily member implicated in development of the nervous system and organ epithelia. Mol Cell Neurosci (2003) 22: 62-74.

Bentley L, Nakabayashi K, Monk D, Beechey C, Peters J, Birjandi Z, Khayat FE, Preece MA, Stanier P, Scherer SW and Moore GE.  The imprinted region on chromosome 7q32 extends to the carboxypeptidase A gene cluster: An imprinted candidate for SRS. J Med Genet. (2003) 40: 249-256.

Scherer SW, ….., Moore GE, et al. Human Chromosome 7: DNA Sequence and Biology. Science (2003) 300: 767-772.

Murrel A, Heeson S, Cooper WN, Douglas E, Apostolidou S, Moore GE, Maher ER. and Reik W. An association between variants in the IGF2 gene and Beckwith-Wiedemann syndrome: ineteraction between genotype and epigenotype. Hum Mol Genet (2004) 13: 247-255.

Nakabayashi K, Makino  S, Minagawa S, Smith AC, Bamforth JS, Stanier P, Preece M, Parker-KatiraeeL, Paton T, Oshimura M, Mill P, Yoshikawa  Y, Hui C-C, Monk D, Moore GE, and Scherer SW.  Genomic imprinting of PPP1R9A encoding neurabin I in skeletal muscle and extra-embryonic tissues. J Med Genet (2004) 8: 601-608.