Hindawi Dermatology Research and Practice Volume 2018, Article ID 1284568, 9 pages https://doi.org/10.1155/2018/1284568

Research Article Whole Genome Sequencing in an Acrodermatitis Enteropathica Family from the Middle East

Faisel Abu-Duhier,1 Vivetha Pooranachandran,2 Andrew J. G. McDonagh,3 Andrew G. Messenger,4 Johnathan Cooper-Knock,2 Youssef Bakri,5 Paul R. Heath ,2 and Rachid Tazi-Ahnini 4,6

1 Prince Fahd Bin Sultan Research Chair, Department of Medical Lab Technology, Faculty of Applied Medical Science, Prince Fahd Research Chair, University of Tabuk, Tabuk, Saudi Arabia 2Department of Neuroscience, SITraN, Te Medical School, University of Shefeld, Shefeld S10 2RX, UK 3Department of Dermatology, Royal Hallamshire Hospital, Shefeld S10 2JF, UK 4Department of Infection, Immunity and Cardiovascular Disease, Te Medical School, University of Shefeld, Shefeld S10 2RX, UK 5Biology Department, Faculty of Science, University Mohammed V Rabat, Rabat, Morocco 6Laboratory of Medical Biotechnology (MedBiotech), Rabat Medical School and Pharmacy, University Mohammed V Rabat, Rabat, Morocco

Correspondence should be addressed to Rachid Tazi-Ahnini; [email protected]

Received 4 April 2018; Revised 28 June 2018; Accepted 26 July 2018; Published 7 August 2018

Academic Editor: Gavin P. Robertson

Copyright © 2018 Faisel Abu-Duhier et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

We report a family from Tabuk, Saudi Arabia, previously screened for Acrodermatitis Enteropathica (AE), in which two siblings presented with typical features of acral dermatitis and a pustular eruption but difering severity. Afected members of our family carry a rare genetic variant, p.Gly512Trp in the SLC39A4 which encodes a zinc transporter; disease is thought to result from zinc defciency. Similar mutations have been reported previously; however, the variable severity within cases carrying the p.Gly512Trp variant and in AE overall led us to hypothesise that additional genetic modifers may be contributing to the disease phenotype. Terefore whole genome sequencing was carried out in fve family members, for whom material was available to search for additional modifers of AE; this included one individual with clinically diagnosed AE. We confrmed that the p.Gly512Trp change in SLC39A4 was the only candidate homozygous change which was sufciently rare (ExAC allele frequency 1.178e-05) and predicted deleterious (CADD score 35) to be attributable as a fully penetrant cause of AE. To identify other which may carry relevant genetic variation, we reviewed the relevant literature and databases including Consortium, GeneMANIA, GeneCards, and MalaCards to identify zinc transporter genes and possible interacting partners. Te afected individual carried variants in RECQL4 and GPAA1 genes with ExAC allele frequency <0.01 and CADD score >10. p.Gly512Trp is highly likely to be the pathogenic variant in this family. Tis variant was previously detected in a Tunisian proband with perfect genotype-phenotype segregation suggestive of pathogenicity. Further research is required in this area due to small sample size, but attention should be given to RECQL4 and GPAA1 to understand their role in the skin disease.

1. Introduction dermatitis, growth retardation, and behavioural changes. Although AE has been attributed to fully penetrant homozy- Acrodermatitis Enteropathica (AE) is a rare inherited gous mutations in the SLC39A4 gene, severity is variable metabolic condition that afects zinc absorption and inher- even within individual families [3] suggesting that additional itance is ofen seen in an autosomal recessive pattern [1]. geneticmodifersmayplayarole. Te frequency of inherited cases is estimated at 1:500,000 Measurement of plasma zinc is the commonest method of individuals with no obvious correlation in race or gender identifying patients with AE, and reduced levels <60 ug/dL [2]. Te manifestations of AE include alopecia, diarrhoea, were noted in patients B03 and B06 in the original study 2 Dermatology Research and Practice

Figure 1: Pedigree analysis to demonstrate the family of AE and the sequenced individuals for this study. Genome sequence of the following samples: A01, A02, B03, AB01, and AB02. Only B03 and B06 are afected (one male sibling was also afected, but he died). Note. Shading in blue displays the individuals afected by the disease. which were correlated with their typical AE symptoms. now been evaluated. In this study, we aimed to identify However, limitations of zinc testing have been highlighted pathogenic and relevant modifer genes associated with by Garza-Rodriguez et al. (2015) [4] who reported a rare familial AE. On this occasion the proband B03, his parents case of AE presenting with two novel missense mutations A01 and A02, and unafected family members AB01 and AB02 of the SLC39A4 gene associated with periorifcial and acral were examined by whole genome sequencing. Two additional dermatitis only and with normal plasma levels of zinc. family members were utilised as population controls in order Te family in this study was previously screened and to eliminate shared, but nonrelevant alleles. analysed by Abu-Duhier et al. 2017 [3] to identify the pathogenic mutation. In this analysis DNA samples obtained 2. Materials and Methods from eleven individuals were amplifed by polymerase chain reaction and Sanger sequenced. Tis comprised both parents Ethical approval was obtained from both University of Tabuk and all the siblings including the afected individuals with and University of Shefeld, and written informed consent their respective partners. A homozygous alternate allele in was obtained to use genetic data for research purposes. 8 at position 145638714 was identifed in the Blood samples were available from 5 members of the fam- gene SLC39A4 (solute carrier 39 member 4) which is a ily with hereditary AE and were initially sequenced (as zinc transporter. Tis mutation was present only in the discussed above), followed by whole genome sequencing. two afected members B03 and B06 presenting with the AE Te individuals studied were the afected child B03, his phenotype(Figure1).Temutationgaverisetoachange parents A01 and A02, and two unrelated family members from the neutral aliphatic glycine to the highly AB01 and AB02 who were included as individuals from the hydrophobic aromatic tryptophan, with genotype c.1534G>T same population background (Figure 1). Both lanes of three at position p.Gly512Trp. Although there was strong evidence HiSeq2500RapidRunwereusedtomultiplexandsequence that this mutation is the cause of AE in this family, sever- the fve samples. To produce fastq fles, bcl2fastq version ity was variable suggesting that other modifers may be 1.8.4 was used with adapter trimming further modifed to present. Moreover, because sequencing was limited to a small accept a single mismatch in the index sequence. BWA ALN number of genes, it is possible that alternative homozygous version 0.7.5a was used to map reads by lane to the human pathogenic change in a zinc transporter gene may have been reference genome hg19. Picard version 1.101 was used to missed. Others have reported the p.Gly512Trp change in a mark duplicate reads followed by realignment around InDels patient with AE [1], but the role of genetic modifers has not using GATK version 2.6-5-gba531bd. Picard was then used previously been examined. to merge lane-level bam fles, with additional marking of To exhaustively defne the genetic basis of AE in our duplicates and realignment around InDels on the complete patients, the whole genome of available family members has bam fle. GATK HaplotypeCaller was used to call variants. Dermatology Research and Practice 3

Table 1: “R” sofware was used to identify the deleterious zinc transporter gene in B03. An EXAC frequency of <0.01 and CADD score of >10 were accepted. Excluding synonymous variants, at position Chr8:145638714 of the SLC39A4 gene, c.G1534T and p.G512W,portrayed as a rare mutation in comparison to other identifed genes and genetic variants. Note. Please see Sup Materials 2 for complete list of zinc transporter genes derived by “R” sofware for B03.

Gene Chromosome Start-End Exon Nucleotide substitution substitution EXAC Frequency CADD Score SLC39A4 Chr8 145638714 10 c.G1534T p.G512W 1.178e-05 35

Te minimal calling quality was accepted at 1, allowing 10 genes which are known to interact with SLC39A4, we used alternative haplotypes. GATK was further used to calculate GeneMANIA (https://genemania.org) to identify relevant coverage statistics. protein-protein interactions, coexpressed genes, and genes Variant calling fles of the fve individuals were automat- of related function based on transcription and phenotypic ically annotated using wANNOVAR to provide information screening profles. Te interaction databases used by Gen- including genomic annotation, frequency of variant observed eMANIA included BioGRID and Pathway Commons, both in controls, and CADD scores. “R” sofware was used to anal- including primary research studies. Databases GeneCards yse the annotated variants. Relevant literature was reviewed and MalaCards were then used to obtain information on the and GO consortium data used to identify all zinc transporter identifed genes (Sup Materials 3). Te interacting genes were genes including SLC39A4. Variants in exonic regions of zinc screened for rare deleterious variants which may have nega- transport genes were fltered by ExAC frequency and CADD tively impacted upon the function of SLC39A4 and thus the score using “R” to identify rare, predicted deleterious variants AE phenotype in our patient. With specifc focus on exonic present in a homozygous form (see sup materials). An ExAC regions of the genes and nonsynonymous mutations, “R” frequency <0.01 and CADD score >10 were employed to analysis demonstrated two genes that were within acceptable suggest a deleterious variant. (ExAC allele frequency suggests ExAC allele frequency and CADD score range (<0.01, >10): the relative frequency of an allele at a genetic locus in GPAA1 and RECQL4, suggestive of signifcant interaction a population [5]; CADD score helps measure the level of withSLC39A4indevelopingthediseasephenotypeinB03. toxicity of a genetic variant such that a value of 10 defnes the variant to be within 10% of a damaging variant in the .) Te analysis was repeated in the other sequenced 4. Discussion individuals to determine whether the toxic mutation was present in relatives. AE appears to occur mainly in France and Tunisia. Te To extend the analysis beyond zinc transporter genes, p.Gly512Trp variant was previously identifed in a Tunisian GeneMANIA was used to identify genetic interactions of family [1]. All published mutations in the human SLC39A4 SLC39A4. Interacting partners were screened for poten- gene have been collated (Table 2). In the majority of cases, tial rare deleterious variants although the requirement for pathogenic variants were present in a homozygous or com- homozygosity was relaxed to include heterozygous variants. pound heterozygous state in AE. Families identifed across a number of countries with AE have been shown to carry over 30 diferent mutations in SLC39A4, including deletions, 3. Results nonsense, missense, and splice-site alterations [6]. Schmitt et al. [1] reported mutations including p.Gly512Trp and 3.1. Only the Homozygous p.Gly512Trp SLC39A4 Variant Is p.549delLeu afecting amino acids that were highly conserved Sufciently Rare and Deleterious to Cause AE in Our Pedigree. between a number of species. Variants p.Gly512Trp and Te identifed deleterious SLC39A4 variant p.Gly512Trp p.549delLeu were detected in homozygous state in Tunisian (with ExAC frequency of 1.178e-05 and CADD score of 35) and Swedish probands, respectively, and the toxic mutation was present in homozygous form in B03 (Table 1). As would was not identifed in any of 164 control from be expected the same p.Gly512Trp change was found to be North Africa included in their study. Moreover, there was present in an heterozygous form in both parents, A01 and perfect segregation between the genotype and phenotype A02, but was not found in the unafected relatives. Te in the pedigree suggesting a high likelihood for AE to be remainder of identifed variants in SLC39A4 and all other associated with SLC39A4 gene. As previously described by screened zinc transport genes demonstrated either a low Abu-Duhier et al. [3], the mutation was located in a putative CADD score or a high ExAC frequency implying a benign transmembrane domain and likely to alter zinc absorption or uncertain signifcance (Sup Materials 2). by reducing transcription/ translation of SLC39A4. Heterozy- gous individuals tend to be asymptomatic carriers rather than 3.2. Screening SLC39A4 Interacting Partners Identifed Rare manifesting the disease phenotype [7]. A signifcant feature of Deleterious Variants in Candidate Modifer Genes. Te AE overall and our family in particular is variable phenotype patient B03 sufered a particularly severe form of AE even between individuals with the same genetic change in and therefore is a good candidate to identify additional SLC39A4 [3]. It is anticipated the AE phenotype could be deleterious genetic modifers of AE. To identify candidate dependent on either modifer genes or an unknown putative 4 Dermatology Research and Practice Study Kury et al. 2003 Kury et al. 2003 Kury et al. 2002 Kury et al. 2002 Kury, et al. 2003 Kury et al., 2002 Wang et al. 2002 Wang et al. 2002 Kharf et al. 2005 Nakano et al. 2003 Nakano et al. 2003 Schmitt et al. 2009 Coromilas et al. 2011 tion of interest that has been (i) Missense (ii) Japanese (iii) Compound heterozygous (iv) Abolishes restriction site for Faul (i) Missense (ii) France (iii) Compound Heterozygous (iv) Deletion in one alleleof – gene failed expression (i) Missense (ii) Various countries (iii) Possibly compound heterozygous (iv) Various amino acid change (i) Missense (ii) May afect protein conformation dueloss to of disulfde bond (iii) Tunisia (iv) Homozygous (i) Homozygous (ii) Frameshif (i) Nonsense (ii) Homozygous (iii) Tunisia (iv) Lacks putative zinc binding site (i) France (ii) Compound heterozygous/homozygous (iii) Possibly altering transcripts through mis-splicing (i) Nonsense (ii) Homozygous (iii) France (iv) Appearance of premature stop codon (i) Missense (ii) Heterozygous (iii) Caucasian (i) Missense (ii) France (iii) Homozygous (i) Missense (ii) reduces Vmax/alter protein folding (iii) France + Austria (iv) Compound heterozygous/homozygous . Likely Pathogenic Pathogenic Pathogenic Pathogenic /Pathogenic Likely Benign Likely/benign Likely pathogenic Uncertain Signifcance p.Leu48X (possibly) (possibly) p.Val171Leu Uncertain Signifcance p.Pro84Leu p.Arg95Cys p.Cys62Arg Uncertain Signifcance p.Asn106Lys p.Glu284Lys p.Pro200Leu p.Gln398fsX18 Donor splice site error Acceptor splice site error > G > T > A > T > T > C > A > T > G c.511G c.251C c.184T c.143T c.318C c.283C c.599C c.1191insC c.850G c.475-2A > A c.192+19G Mutation (Nucleotide) Infuence on Amino Acid Clinical Signifcance Other fndings Exon Exon 6 Exon 5 Exon 1 Exon 1 Intron 1 Exon 2 Exon 2 Intron 2 Exon 2 Exon 3 Exon 3 identifed in a Tunisian family. Note. Some reports failed to provide adequate information to complete the table Table 2: Modifed table from Schmitt et al. (2009) on genetic mutations identifed up to date with AE associated SLC39A4 gene. Outlined in bold is the muta Dermatology Research and Practice 5 Study Li et al. 2010 Kury et al. 2003 Kury et al. 2002 Kury et al. 2002 Wang et al. 2002 Wang et al. 2002 Wang et al. 2002 Wang et al. 2002 Wang et al. 2002 Mefah et al. 2006 Nakano et al. 2003 Nakano et al. 2003 Santiago et al. 2011 Schmitt et al. 2009 Schmitt et al. 2009 (i) Frameshif (ii) France (iii) Compound Heterozygous (iv) Alter protein function (i) Missense (ii) Homozygous (iii) France (i) Missense (ii) Heterozygous (iii) Caucasian (i) Deletion (ii) Spanish (iii) Truncated protein (iv) Heterozygous (i) Frameshif (ii) Premature termination codon (iii) Heterozygous (iv) Japan (i) Nonsense (ii) Truncated protein (iii) Tunisia (iv) Homozygous (i) Missense (ii) Homozygous (iii) Substitution of highly conserved amino acid (iv) Japan (i) Cellular mislocalization (ii) Missense (iii) Egypt (iv) Homozygous (i) Reduced protein levels (ii) Missense (iv) Egypt (iv) Homozygous (i) Reduced protein levels (ii) Missense (iii) France (iv) Homozygous (i) Homozygous (ii) France (i) Missense (ii) Homozygous Likely/ Benign Unknown Unknown (i) Missense Unknown (i) Frameshif Unknown (i) Missense Pathogenic Pathogenic Pathogenic Likely/benign Likely Pathogenic Likely Pathogenic Likely Pathogenic Uncertain signifcance Table 2: Continued. (possibly) p.Gln211X p.Tr381Ala Uncertain Signifcance p.Arg251Trp p.Glu284Lys p.Leu372Pro p.Gly374Arg p.Gln303His p.Cys309Tyr p.Gly330Asp p.Leu372Arg p.Tr357AlafsX10 p.Leu256SerfsX16 Likely Pathogenic p.Ser214ArgfsX30 p.Ser324ArgfsX24 Acceptor splice site error > G > C > G > A > A > C > G > T > T 971del 1017ins53 642ins10 AGTC c.751C c.631C c.766delC c.1115T c.1115T c.850G c.926G > A c.989G c.909G c.1141A > A c.1120G c.1150-2A c.968 c.641 c.1016 Mutation (Nucleotide) Infuence on Amino Acid Clinical Signifcance Other fndings Exon Exon 3 Exon 3 Exon 4 Exon 4 Exon 5 Exon 5 Exon 5 Exon 5 Exon 6 Exon 6 Exon 6 Exon 6 Exon 6 Exon 6 Intron 6 6 Dermatology Research and Practice Study Park et al. 2010 Kury et al. 2003 Kury et al. 2002 Kury et al. 2002 Vardi et al. 2009 Wang et al. 2002 Wang et al. 2002 Wang et al. 2008 Kharf et al. 2010 Schmitt et al. 2009 Nakano et al. 2009 Schmittetal.2009 (i) Nonsense (ii) Compound Heterozygote (iii) Austria (iv) Absence of zinc binding site (i) Frameshif (ii) Founder Efect (iii) Tunisian (iv) Homozygous (i) Reduced protein levels (ii) Missense (iii) Homozygous (iv) Jordan (i) Reduces Vmax (ii) Missense (iii) France (iv) Homozygous (i) Missense (ii) Tunisia (i) Deletion (ii) Sweden (iii) Homozygous (iv) Afect amino acids (i) Missense (ii) Tunisia (iii) Homozygous (iv) Afect amino acids Likely/ Unknown Stop Unknown (i) Frameshif Unknown (i) Frameshif pathogenic Pathogenic Pathogenic Pathogenic retardation/X-Linked Pathogenic for mental Uncertain Signifcance Table 2: Continued. X 1 0 4 p r T . (possibly) p.Gly595Val Uncertain signifcance p.549delLeu p.Glu480Sto p.Leu410Pro Uncertain Signifcance (i) Missense p.Gly526Arg p.Gly630Arg p.Gly512Trp p.Ala408fsX481 p.Trp411ArgfsX7 Uncertain signifcance Acceptor splice site error > T > T > C > Ap > A 1648delTGC 1227delCCGGG c.1223delC > C c.1229T > T c.1438G c.1784G c.1576G c.1888G c.1203G c.1534G > C c.1287+2T c.1646 c.1223 Mutation (Nucleotide) Infuence on Amino Acid Clinical Signifcance Other fndings 147411+delAGACTGAGCCCAGG p.Arg488SerfsX2 Exon Exon 7 Exon 7 Exon 7 Exon 7 Intron 7 Exon 9 Exon 9Exon 10 c.1462 Exon 10 Exon 11 Exon 11 Exon 12 Dermatology Research and Practice 7 Tymus. epithelium. Tissue Expression epithelium and breast entify the function, disorder, blood, testis and breast. Overexpressed in Nasal Overexpressed in heart. Ubiquitously expressed. fetal gut and CD8 Tcells. Esophagus, stomach and Intestine, Ovary and Testis. Overexpressed in pancreas. Mostly Lungs and intestine. Overexpressed in Testis and Nervous system, skin, lungs. Lungs, Skin, Nervous system. Skin. Overexpressed in whole pancreas. Overexpressed in oral Overexpressed in small intestine, N/A N/A slow growth, ketoacidosis. Signs/Symptoms insulin-responsive hyperglycemia and growth retardation. Growth retardation, Infections, persistent urinary incontinence, Clef lip, myelocystocele, leukocytosis, mental and Lactic acidosis, infection, Fusion of bones (neonates), hydrocephalus and lipomas. immune-system dysfunction, alopecia, diarrhea, dermatitis. missing/malformed kneecaps. Function membrane. segregation. cytochrome. the intestine. sulphur protein to GDP-D-mannose. Catalyzes the reactions modulate chromosome Mediates the transfer of Regulates epithelial and stranded DNAs and may Encodes for a ZIP family. neuronal morphogenesis. electron from Rieske iron epimerase and reductase in Required for zinc uptake in Participates in cellular RNA processing and degradation. Linksproteinstocellsurface DNA unwind double N/A N/A Disease defciency defciency, type II Neural tube defects. Leukocyte adhesion Tick-Borne Encephalitis. Baller-Gerold Syndrome. . Mitochondrial complex III Acrodermatitis enteropathica Protein Member 4 Description Tissue Specifc Cytochrome C1 Anchor Attachment RecQ Like 4 Exosome Component 4 Solute Carrier Family 39 Glycosylphosphatidylinositol Scribbled Planar Cell Polarity Transplantation Antigen P35B Gene SLC39A4 TSTA3 SCRIB GPAA1 CYC1 RECQL4 EXOSC4 and tissue expression. Note. Te documented information only includes the main signs, symptoms, and tissue expression. Table 3: GeneMANIA presented the interacting partners with SLC39A4 gene. Te databases GeneCards and MalaCards and the literature were reviewed to id 8 Dermatology Research and Practice

Table 4: GPAA1 and RECQL4 genes both demonstrated a likelihood of being modifer genes of SLC39A4 in developing AE in B03. Note. Please see Sup Materials 3 for “R” sofware version of complete list of interacting partners with SLC39A4 gene for B03.

Gene Chromosome Start-End Exon Nucleotide substitution Protein substitution EXAC Frequency CADD Score GPAA1 Chr8 145140564 11 c.G1540A p.A514T 0.0053 15.04 RECQL4 Chr8 145740364 9 c.C1576T p.L526F 0.0007 16.37

AE gene. Hence we have screened for additional genetic Consent modifers of SLC39A4 function within an individual with a particularly severe AE phenotype [3]. Te participants gave their consent afer they received verbal Ourdatasuggeststhatthep.Gly512Trpchangeisthe and written explanation including patient information. Indi- likely cause of disease in our family because it is the only vidual consent has been signed by each individual. homozygous, rare, and predicted deleterious change within a zinc transporter gene which is present within the afected patient and absent (or heterozygous) in unafected family Disclosure members. In addition we identifed two rare and predicted Faisal Abu-Duhier is the Dean of the Faculty of Applied deleterious genetic variants in SLC39A4 interacting partners Medical Science; Andrew Messenger and Andrew McDonagh whichmayhavearoleinthedevelopmentoftheparticularly are Consultant Dermatologists at Shefeld Teaching Hospi- severe AE phenotype in our patient: RECQL4 and GPAA1 tals; Youssef Bakri is the Head of the Biology Department, (Table 4). Interestingly Nistor et al. [8] reported that the Faculty of Sciences, Rabat; and Vivetha Pooranachandran, complete clinical triad of features in AE was only documented Paul Heath, Johnathon Cooper-Knock, and Rachid Tazi- in 20% of patients, questioning if only one gene is responsible Ahnini are Research Scientists at the University of Shefeld. and if modifer genes could be involved in the variable clinical Te authors authorised the publication of this manuscript. characteristics. Patients with mutation in the RECQL4 gene have several characteristic features similar to AE; for example, Bernstein et al. [9] reported a RECQ disorder: Bloom syn- Conflicts of Interest drome, a rare autosomal recessive condition, which presents with mental retardation, immunodefciency, male infertility, Te authors declare that they have no conficts of interest. and increased chance of cancer. Similar features occur in some patients with AE, in the form of immunodefciency, mental retardation, and infertility. In addition, Mann et al. Authors’ Contributions [10] identifed distinctive skin abnormalities in a mutant RECQL4 mouse model. Although the GPAA1 gene is known Rachid Tazi-Ahnini and Faisel Abu-Duhier were the prin- to be expressed in skin (Table 3), no disorder has been cipal investigators. Faisel Abu-Duhier was responsible for identifed to date that has been directly afected by GPAA1 the acquisition of clinical materials. Faisel Abu-Duhier, Paul genetic mutation, although rare cases have been reported R. Heath, and Rachid Tazi-Ahnini were responsible for the in which GPAA1 gene amplifcation and RNA and protein concept and design of the study. Vivetha Pooranachandran, overexpression occurred in hepatocellular carcinoma [11]. Rachid Tazi-Ahnini, and Paul R. Heath were responsible for Further research is required to explore a putative link between the analysis and interpretation of data. Vivetha Pooranachan- GPAA1 and RECQL4 mutations and AE. dran and Johnathan Cooper-Knock were involved in design- Our data supports pathogenicity of the p.Gly512Trp ing sofware for genetic analysis. Andrew J. G. McDonagh, change in SLC39A4, but in addition we have identifed poten- Andrew G. Messenger, Youssef Bakri, and Rachid Tazi- tially deleterious variants in SLC39A4 interacting partners Ahnini drafed the manuscript. All authors participated in which may be important to disease pathogenesis and are the study design, critical revision, and fnal approval of potential therapeutic targets. the manuscript. Paul R. Heath and Rachid Tazi-Ahnini contributed equally to this work. Data Availability Acknowledgments Tedatausedtosupportthefndingsofthisstudyare available from the corresponding author upon request. Tis work was funded by University of Tabuk Prince Fahd Bin Sultan Research Chair in collaboration with University Ethical Approval of Shefeld. Research contact grant number is R/130649- 11-1. Te authors are grateful to the family from Tabuk for Tis work complies with ethical standards. Te study was participating in this study. Te authors also thank Shefeld approved by University of Tabuk and University of Shefeld Diagnostic Genetics Service, Shefeld Children’s NHS Trust, ethical committees. for genome sequencing. Dermatology Research and Practice 9

Supplementary Materials Supplementary materials include codes derived for “R” sof- ware analysis of the genome data, code used to identify the mutated zinc transporter genes in B03 sample, and code used to identify interacting partners with SLC39A4 gene in causing the disease in B03. (Supplementary Materials)

References

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