PDF hosted at the Radboud Repository of the Radboud University Nijmegen The following full text is a publisher's version. For additional information about this publication click this link. http://hdl.handle.net/2066/22098 Please be advised that this information was generated on 2021-04-06 and may be subject to change. http://hdl.handle.net/2066/22098 Hum Genet ( 1995) 95:157-160 Tuija Sulisalo • Ineke van der Bürgt • David L. Rimoin Jaques Bonaventure • David Sillence • John B. Campbell David Chitayat • Charles I. Scott • Albert de la Chapelle Pertti Sistonen • Ilkka Kaitila Genetic homogeneity of cartilage-1 Received: 8 July 1994 Abstract Cartilage-hair hypoplasia (CHH) is an ¿utoso- mal recessive metaphyseal chondrodysplasia character­ ized by short stature and hypoplasia of the hair. Associ­ ated pleiotropic features include deficient erythrogenesis, impaired T-cell mediated immunity, Hirschsprung’s dis­ ease, and an increased risk of malignancies. CHH is most prevalent among the Old Order Amish in the United States and among the Finns, but sporadic families have been described among many other populations. We have previously mapped the gene for CHH to the short arm of chromosome 9 in Finnish and Amish families. The CHH locus resides close to D9S163 within an interval of 1.5cM flanked by D9S165 and D9S50. In order to investigate the genetic homogeneity of CHH in various populations, we studied nine families with no genealogical connections to T. Sulisalo (CE3) * A. de la Chapelle • I. Kaitila Department of Medical Genetics, P.O. Box 21, FIN-00014 University of Helsinki, Helsinki, Finland I. van der Bürgt Department of Human Genetics, University Hospital Nijmegen, Nijmegen, The Netherlands D. L. Rim oin Cedars-Sinai Medical Center, Los Angeles, USA J. Bonaventure CNRS, Hospital Necker, Paris, France D. Sillence The Children’s Hospital, Sidney, Australia J. B. Campbell Arnold Palmer Hospital for Children & Women, Orlando, USA D. Chitayat The Hospital for Sick Children, Toronto, Canada C. I. Scott Alfred I. duPont Institute, Delaware, USA P. Sistonen Finnish Red Cross Blood Transfusion Service, Helsinki, Finland I. Kaitila Department of Medical Genetics, Helsinki University Central Hospital, Helsinki, Finland © Springer-Verlag 1995 hypoplasia either Amish or Finns. No recombinants were detected be­ tween the CHH gene and any of the three closest marker loci studied, suggesting that CHH in these families results from mutation(s) at the same locus as in the Amish and Finnish families. Introduction Cartilage-hair hypoplasia or McKusick type metaphyseal chondrodysplasia (M IM No. 250250) is an autosomal re­ cessive disorder resulting in short-limbed short stature and poor hair growth (McKusick et al. 1965; Mákitie and Kaitila 1993). Associated pleiotropic features include de­ ficient erythropoiesis, impaired T-cell mediated immunity, intestinal problems such as Hirschsprung’s disease, and an increased risk of malignancies. Although the pathogenesis of CHH is unknown, defective proliferation of T- and B- lymphocytes and fibroblasts, and defective erythrogenesis suggest a more generalized defect in cellular proliferation. CHH was first described among the Old Order Amish in the United States (McKusick et al. 1965) and was later also shown to be a relatively common cause of short stature among the Finns (Perheentupa 1972). Gene frequencies of 0.05 and 0.0066 have been estimated in these populations, respectively (McKusick et al. 1965; Mákitie 1992). Data on gene frequencies among other populations are not avail­ able, but sporadic families have been described among many populations, including the Dutch, Polish, French, Germans, Danes, Algerians, Italians, Spanish, non-Amish Americans, and the Mexicans (reviewed in Makitie 1992). We have previously assigned the gene for CHH to chromosome 9 by linkage in Finnish and Amish families (Sulisalo et al. 1993, 1994a). The CHH gene has been mapped to an area of approximately 1.5cM flanked by D9S165 and D9S50, and is close to D9S163 (Sulisalo et al. 1994b). No recombinations have been detected between the CHH gene and D9S163, and a statistically significant linkage disequilibrium was observed between these loci in both Amish and Finnish families (Sulisalo et al 1994a, b). Luria-Delbrück method-based calculations (Hastbacka et 158 D9S165 41 D9S163 4 6 D9S50 41 17 63 13 1 17 63 13 4 7 43 4 3 77 63 33 17 66 13 71 46 43 2 77 46 4 2 1 1 66 3 3 71 66 23 1 1 66 33 77 46 34 3 77 43 33 77 63 4 3 77 63 4 3 77 63 4 3 4 7 34 3 3 5 46 34 13 7 5 44 34 57 46 41 57 46 41 77 4 3 3 3 77 38 36 36 31 42 85 63 27 35 33 4 7 65 33 37 77 43 33 8 I r^ \ 7 i u v J 66 53 71 77 36 46 33 35 16 34 3 5 67 4 3 53 66 4 3 53 67 33 33 77 46 13 10 7 4 66 13 74 36 13 57 66 31 77 66 15 47 46 55 77 4 3 33 12 7 6 33 31 16 43 51 ND ND ND 77 34 34 ND ND ND ND ND ND 14 17 36 13 ND ND ND 15 57 24 23 ND ND ND ND ND ND 47 4 6 13 ND ND ND 159 4 Fig. 1 Pedigrees of the families studied. The haplotypes for D9S165, D9S163, and D9S50 are shown for all individuals stud­ ied. The phase could not be determined for the marker alleles in all cases. Square Male, circle female, open symbol unaffected, filled symbol affected, all symbols with diagonal deceased, ND not de­ termined T able 1 Two-point tod scores between the C H H gene and three nearest marker loci in nine C H H fam ilies for w hich neither Finnish nor A m ish ancestors are know n al. 1992; Lehesjoki et al. 1993) suggest an approximate distance of 0.3 cM between C H H and D9S163 (Sulisalo et a l 1994b). Here, we have studied nine C H H families for which neither Amish nor Finnish ancestors are known. In addi­ tion, seven sporadic cases were studied. These families originate from the Netherlands, Poland, France, Croatia, the United States, and Canada. Materials and methods Patients The pedigrees o f the families ¿ire shown in Fig. 1. Families 1 and 2 come from the Netherlands, fam ily 3 from Poland, family 4 from France, fam ily 5 from Australia (originally from Croatia), families 6-8 and 10-16 from the United States, and family 9 from Canada. Diagnosis was performed by Lv.d.B. in families 1-3 (van der Burgt et al. 1991), by J.B. in family 4, by D.S. in family 5, by D .L.R, in families 6-7 and 10-15, by C.I.S. in family 8, by D .C . in family 9, and by J.B.C. in family 16. The diagnostic criteria are short-limbed short stature, generalized laxity of joint ligaments, and metaphyseal 11 tiring and irregularities o f the growth plates in childhood radiographs. Poor hair growth is used only as a positive criterion (M cKusick et al. 1965; Miikitie and Kaitila 1993). D N A samples D N A was extracted from blood or lymphoblastoid cell lines ac cording to standard methods. Polymerase chain reaction and detection of microsatellite markers The polymerase chain reaction protocols for the detection of mi- crosatellite markers have been described previously (Sulisalo et al, 1993). Microsatellite markers detecting D9S165, D9S163* and D9S50 were studied (Weissenbach et al. 1992; W ilkie et al. 1992). Linkage analysis Pairwise linkage analysis was performed using the M L IN K pro­ gram of the L IN K A G E package (Lathrop et al. 1984). M ultipoint analysis against a fixed marker map of three loci, D9S165, D9S163, and D9S50, was carried out using the L1NKM AP pro­ gram and assuming a constant female/male sex difference ratio of 2.75 and distances as previously reported (Sulisalo et al. 1994b). The C H H gene was considered fully penetrant and the frequency was set to 0.001. The marker allele frequencies were calculated from the normal chromosomes of 66 Finnish CH H families (Sulisalo et al. 1994b), Heterogeneity was tested using the A-test as implemented in program H O M O G (Ott 1991). Results and discussion Locus e 0.0 0.001 0.01 0.05 0.1 0.2 0.3 0.4 D 9 S I6 5 2.20 2.20 2,13 1.85 1.51 0.93 0.45 0.12 D9S163 3.23 3.22 3.14 2.75 2.28 1.38 0.64 0.16 D9S50 2.63 2.62 2.55 2.23 1.84 1.13 0.55 0.15 Nine potentially informative families were included in ifi vadous populations. the linkage analysis (families 1-9). The distribution of marker alleles in these families is shown in Fig. 1, The two-point lod scores between the C H H gene and three closest marker loci are shown in Table 1. No recombinations were detected between the C H H gene and markers in any of the informative meioses, suggesting genetic homogeneity of C H H in various populations. The highest lod score of 3.23 at a zero recombination fraction was detected between the C H H gene and D9S163. The m axim um lod scores for D9S165 and D9S50 reached 2 .2 0 and 2.63, respectively. The maximum m ultipoint lod score of 4.10 was obtained at D 9 S 163. No evidence o f heterogeneity was found (H O ­ M O G ), although the sample size was limited and all meioses were not fully infoi*mative for all markers stud­ ied. The genetic homogeneity o f C H H validates the use of closely linked markers for prenatal diagnosis in families from various populations. We have previously observed a statistically significant association between the C H H gene and alleles at D9S163 in both Finnish and A m ish families (Sulisalo et al. 1994a, b). In Finnish families, C H H is associated with allele 3, and in Am ish families, with allele 6 . O f the 32 C H H chro­ mosomes studied here, 14 carried allele 6 , 10 carried al­ lele 3, seven carried allele 4, and one carried allele 2 at D9S163. Alleles 3, 4, and 6 are common in the normal chromosomes of the Finnish and A m ish C H H families, and in the Centre d ’Etude du Polymorphisme H um ain (CEPH) reference families (frequencies vary from 0.2 to 0.4; data not shown). A m ong the Finnish chromosomes, allele 3 is the most com m on (41% ) and alleles 4 (33%) and 6 (19%) the second and third most common (Sulisalo et a l 1994b). A m ong the Am ish and the CEPH families, however, the most com m on allele is 4 (43% in both se­ ries) and alleles 3 and 6 are the second and third most common (Sulisalo et al. 1994a; C EPH data not shown). As C H H is associated with a common allele at D9S163 among both the Finns and the Am ish, no definitive con­ clusions can be drawn about the origin o f the C H H muta­ tions in the families studied here. However, as most of the latter patients are heterozygous for alleles at D9S163, it can be concluded that either there are many different C H H mutations and a C H H mutation is a more common occurrence than has been assumed, or only a few muta­ tions exist but recombinations or mutations at markers have changed the original marker alleles. Characterization of the gene will be required to determine whether a single or different C H H mutations are responsible for the disease A cknow ledgem ents W e thank Sophie M arc (Genethon, Paris, France) for .sharing the allele frequency data on D9S163 in CEPH families. M aryann Priore and D aniel C ohn (Cedars-Sinai Medical Center, Los Angeles, U SA ), and Judith Hall (British Colum bia Childrens Hospital, Vancouver, Canada) are acknowledged for collaborative help. This study was financially supported by the March o f Dim es Birth Defects Foundation (#6-FY92-0181), the Sigrid Juselius Foundation, the Academy o f Finland, and the Paulo Foundation. Part o f the study was carried out at the Folkhalsan In ­ stitute of Genetics. References Burgt 1 van der, Haraldsson A , Oosterwijk JC , Essen AJ van, Wee- maes C, Hamel B (1991) Cartilage hair hypoplasia, metaphy­ seal chondrodysplasia type M cKusick: description of seven pa­ tients and review o f the literature. 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