Received 24 May 2008 | Accepted 29 November 2008 | Published 12 December 2008
The first three authors contributed equally to this work.
1The Key Sericultural Laboratory of Agricultural Ministry; College of Biotechnology, Southwest University, Beibei, Chongqing, China; 2Center for Human Molecular Biology & Genetics, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Sichuan, China; 3Department of Ophthalmology, Xihua Hospital, Shanghai Jiaotong University, Shanghai, China; 4Linshui County Hospital, Linshu, Sichuan, China; 5Department of Ophthalmology, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Sichuan, China; 6Department of Neurology, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Sichuan, China
Correspondence to: Dr. Qingyou Xia, the Key Sericultural Laboratory of Agricultural Ministry; College of Biotechnology, Southwest University, Beibei, Chongqing 400715, China; Phone: 86-23-68250099; FAX:86-23-68251128; email: email@example.com
Purpose: To describe the clinical features of and identify a novel mutation in Bardet–Biedl syndrome 7 gene (BBS7) in a Chinese family.
Methods: Nineteen individuals at risk for inheriting Bardet-Biedl syndrome (BBS) in a Chinese family participated in the study. Physical examination was performed and blood was drawn for DNA extraction. Linkage analysis was conducted for all known BBS loci, and mutation screening of BBS7 gene and BBS12 gene was performed.
Results: A Chinese family with inherited BBS was identified. After performing linkage analysis on all 13 known loci, we found the disease phenotype of a Chinese family with BBS linked to a locus where BBS7 and BBS12 genes locate.
Conclusions: This study describes a novel mutation in BBS7 causing BBS in a Chinese family. This is the first report that a mutation in a BBS gene causes BBS in a Chinese population. These results expand the spectrum of human disease associated with mutations of BBS7 since the initial three mutations in BBS7 were first identified in 2003.
Bardet–Biedl syndrome (BBS; OMIM 209900) is an autosomal recessive disorder characterized by pleiotropic defects including obesity, retinal dystrophy, polydactyly, hypogenitalism, learning difficulties, and renal abnormalities [1,2], as well as diseases such as diabetes mellitus, hypertension, and congenital heart disease [1,3,4]. The disorder displays genetic heterogeneity. So far, fourteen genes have been mapped and cloned: including BBS1 (11q13), BBS2 (16q21), BBS3 (3p12-q13), BBS4 (15q22.3), BBS5 (2q31), BBS6 (20p12), BBS7 (4q27), BBS8 (14q32.11), BBS9 (7p14), BBS10 (12q21.2), BBS11 (9q33.1), BBS12 (4q27), BBS13 (17q23), and BBS14 (12q21.3) [2,5-18]. There are still about 25%–50% of BBS families without any mutation in these genes [10,19,20]. In vitro and in vivo studies of the known BBS genes and their products have demonstrated that BBS is a disorder of defective primary ciliary and basal body function, probably affecting both transport and paracrine signals such as the planar cell polarity pathway [2,10,12,21,22]. A relatively high incidence of BBS is found in the mixed Arab populations of Kuwait and in Bedouin tribes throughout the Middle East, most likely due to the high rate of consanguinity in these populations and a founder effect. Three mutations linked to BBS have been previously described in BBS7 (T211I, K237fsX296, and H323R) . However, no BBS gene mutation has been reported in Chinese populations. Here we report for the first time that a novel mutation in BBS7 gene causes BBS in a Chinese family.
Approval from the Institutional Review Board of the Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, China was obtained for this study, and informed consent was obtained from all participants. The study enrolled 19 individuals from a Chinese family who were at risk for inheriting BBS. Physical examination was performed, and blood was drawn for DNA extraction. Individuals with BBS were ascertained on the basis of criteria established elsewhere, including the cardinal features such as retinal dystrophy or other ocular abnormality, obesity, learning difficulties, polydactyly, hypogenitalism (males), renal abnormality, and other clinical features . Measurements were made of height and weight. Photographs were taken of the entire body including the hands, feet, and specific dysmorphic features. Renal function was tested. The eyes were examined with retinal-function testing (color-vision testing, perimetry, and dark-adaption testing) and electroretinographic studies, photography of the fundus, and fluorescein angiography. Urogenital system examination and intelligence level evaluation were also conducted.
DNA was extracted from blood samples, and genetic linkage was assessed using short tandem repeat (STR) markers using established methods [23,24]. The STR markers encompassing known BBS loci including BBS1 (D11S987, D11S4191), BBS2 (D16S415, D16S503), BBS3 (D3S3619, D3S1752), BBS4 (D15S131, D15S205), BBS5 (D2S2330, D2S335), BBS6 (D20S115, D20S186), BBS7/BBS12 (D4S402, D4S427, GATA30B11), BBS8 (D14S68, D14280), BBS9 (D7S516, D7S484), BBS10 (D12S376, D12S326), BBS11 (D9S1811, GGAT11B01), BBS13 (D17S787, D17S944), and BBS14 (D12S326, D12S351) were genotyped and analyzed. BBS7 and BBS12 mutation screening was performed by direct sequencing of PCR-amplified DNA fragments for all exons using established methods . Amplified products were purified using the QIAquik Gel Extraction Kit (Qiagen, Valencia, CA) and sequenced with forward and reverse primers by the BigDye® Terminator v3.1 Cycle Sequencing Kit (ABI Applied Biosystems, Foster City, CA) according to the manufacturers’ instructions.
A Chinese consanguineous BBS family was identified in southwest China. Of the 19 members at risk for inheriting BBS in this family, five were diagnosed with BBS (Figure 1). Two affected patients died of severe fever when they were five years of age, and one affected patient died from an accident. The two living affected patients displayed gross obesity in relation to the average height of adult Chinese men. In addition, these patients exhibited the following features: atypical retinal dystrophy with attenuated vessels, choroidal sclerosis-type change, severe optic disc pallor and severe macular dystrophy (Figure 2), obviously constricted visual fields, severe abnormalities of color vision, reduced rod and cone electroretinograph (ERG) response, refractive errors of −10 (IV:2) and −8.0 (IV:4) diopters, polydactyly of hands and feet, hypogenitalism (small testes and genitalia), learning difficulties, and renal abnormalities. The detailed clinical information for these two affected patients is listed in Table 1. Based on their symptoms, the affected patients were given a diagnosis of BBS [1,2].
The genotyping results showed that the BBS7/BBS12 encompassed STR markers, D4S402, D4S427, and GATA30B11, completely cosegregated with the phenotype in the family (Figure 1). But no linkage was found to any other known BBS loci. No mutation was found in BBS12. Mutation screening in the 19 exons of BBS7 found a 1666 A>G mutation in exon 15, which resulted in an amino acid substitution from serine to arginine at position 556 (Figure 3). This mutation was absent in 100 normal matched controls.
It is clear that BBS is a pleiotropic, autosomal recessive disorder. The cardinal phenotype in this family was identical to most cases described previously [1,2] except that the ocular phenotype of this family showed macular dystrophy instead of a typical retinitis pigmentosa in BBS, further indicating the variant retinal dystrophy in different BBS families. The ocular manifestations of BBS included an early and usually severe rod-cone dystrophy that is detected by ERG and causes central and peripheral visual loss by the second to third decades of life. More than 70% of the affected individuals are legally blind by this time. High myopia associated with choroidal sclerosis-type changes in our family further confirmed the association of a myopia refractive error trend with BBS [19,25].
To date, 13 loci and 14 genes have been mapped and identified since the first BBS gene, MKKS, was identified in 2000 [2,18,25]. CCDC28B modifies the expression of BBS phenotypes in the patients who have mutations in other genes . Mutations in BBS1 constitute the most mutations in the known BBS families (about 23%). Mutations in BBS10, BBS2, and BBS12 account for 10%, 8%, and 5% of all mutations, respectively. Other known BBS genes are rarely mutated in the known BBS families [10,19]. The mutations in these 14 genes probably affect both transport and paracrine signals, such as the planar cell polarity pathway, and cause the pleiotropic defects in affected BBS patients [2,10,12,16,18,20-22,26].
In 2003, Badano, et al.  performed phylogenetic and genomic studies in which they used the human and zebrafish BBS2 peptide sequences to search dbEST and the translation of the draft human genome. They identified a novel gene, BBS2L1. BBS2L1 mutations cause BBS, defining a novel gene for this syndrome, BBS7. BBS1, BBS2, and BBS7 share a distinct substructure, and they may belong to a distinct subfamily of proteins [2,17]. Two transcript variants encoding distinct isoforms have been identified for this gene. The length of isoform 1 is 43 amino acids longer than that of isoform 2. Isoform 2 is ubiquitously expressed, whereas isoform 1 is expressed in retina, lung, liver, testis, ovary, prostate, small intestine, liver, brain, heart, and pancreas. BBS7 may play a role in eye, limb, cardiac, and reproductive system development. Blacque et al.  showed that mutations in the Caenorhabditis elegans bbs-7 and bbs-8 genes cause structural and functional defects in cilia. They further demonstrated that BBS7 and BBS8 are required for the normal localization and motility of the intraflagellar transport (IFT) proteins so the BBS7 protein plays an important role in the assembly as well as function of IFT particle components. These findings indicate that the cardinal and secondary symptoms of BBS may result from cilia . How the mutation in our family caused BBS is not understood as this mutation is located closer to the C-terminal of BBS7 compared with the three mutations reported previously. The initially identified three mutations are located in a predicted β-propeller region of BBS7 that is conserved between BBS2 and BBS7. These findings may indicate that the β-propeller region of BBS7 and the region involving the mutation identified in this study play a critical role in the IFT system. The mutations in these regions probably disturb the localization and motility of the IFT proteins [17,21].
BBS is a rare development disorder, and 14 genes that cause BBS have been identified. However, because of the significant clinical and genetic heterogeneity of BBS, there are still unknown genes involving at least 25%–50% of BBS families [10,15,16,27]. Studies done in more families, especially from different ethnic populations, will be useful to help identify other BBS disease-causing genes and mutations and to better understand the disease mechanism of BBS. Because no BBS mutation has been identified in a large Chinese population, the novel BBS7 mutation reported here will expand the insight into our understanding of BBS.
We thank the patients who participated in our study and their families. We acknowledge the grant support to Z.Y. (The Department of Science and Technology of Sichuan Province 04JY029–045, 05ZQ026–018 and National Natural Science Foundation of China 30671182, 30771220) and to Y.L. (National Natural Science Foundation of China 30771219). Dr. Zhenglin Yang, Center for Human Molecular Biology and Genetics, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, and Dr. Qingyou Xia, the Key Sericultural Laboratory of Agricultural Ministry; College of Biotechnology, Southwest University, are both contribute equally to this research and are considered co-corresponding authors.