Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. mi addresses: Email 1 China 330006, c Child and China Maternal b Provincial 330006, Jiangxi Jiangxi Province, Nanchang, Jiangxi Hospital, of Health Health Reproductive Women’s of Laboratory Key a Wu, Meiling Liu, genesare Xianxian transporters’ study case-control ABC a the title: disease: Running in pregnancy mutations of novel cholestasis withintrahepatic the associated TBA, identifies WES, sequencing Keywords patients. Whole-exome ICP for mutation treatment disease. Ser1342Tyr new ICP gene, provide of ABCC2 architecture and ICP, genetic variants, disease, mutation, the novel one ICP into have genes, insights of which new in transporters’ diagnose patients provide ABC gene the genetic results to ABCC2 to Our compared family Conclusion contribute of HDL ABC may controls. level and in They local TG expression mutations and CHOL, two the ICP DBIL, with in tissue, AST, patients chemical mutation TBA, placental the the no Moreover, of in in value women. average And change pregnant higher slight healthy have chains. than genes a damaging side higher showed in significantly acid structure reported was amino protein were patients first binding mutations ICP for modified ATP-ligand pathogenic ABCC2 Ser1342Tyr novel. of possible and ABCC2 were novel lengths ABCB11 reference, 42 7 ABCB4, bond to which, detected including of compare We genes results, subjected Besides, Results functional prediction were known the group. disorders. patients the to clinical in ICP according located other 151 panels which and four from identified as ICP MAF samples loci with DNA these variants frequencies: and classified Methods genetic allele We variants of (minor patients. genetic Association variants between ICP measures exonic association 151 outcome novel conduct Samples Rare and Jiangxi. DNA, WES. Setting the to disease WES (ICP) acids. pregnancy Design of bile cholestasis approach. total intrahepatic (WES) with sequencing associated variants whole-exome genetic by pathogenic novel the identify To Objectives Abstract 2020 14, August 1 Zheng Liu Xian study case-control of a cholestasis disease: intrahepatic pregnancy with ABC associated the are in genes mutations transporters’ novel the identifies sequencing Whole-exome otiue qal.*orsodn authors. *Corresponding equally. Contributed ffiito o available not Affiliation eta a,JagiPoica aenladCidHat optl acag inx 306 China 330006, Jiangxi Nanchang, Hospital, Health Child and Maternal Provincial Jiangxi Lab, Central eateto bttis inx rvnilMtra n hl elhHsia,Nnhn,Jiangxi Nanchang, Hospital, Health Child and Maternal Provincial Jiangxi Obstetrics, of Department 1 n agZou Yang and , 1 u Lai Hua , a,c a,b,1 isegZheng, Jiusheng B ee oe uain n11IPpatients ICP 151 in mutations novel genes ABC u Lai, Hua 1 iXin Si , 1 a,c,1 a,c,* iigXin, Siming 1 egLi Zeng , agZou, Yang a,c 1 ioZeng Xiao , a,b,* egigLi, Zengming . 1 a 1 ioigZeng, Xiaoming iNie Li , < % eepromdfrsbeun nlss Main analysis. subsequent for performed were 1%) 1 hnLiang Zhen , a,c iuNie, Liju 1 e Wu Mei , a,c hny Liang, Zhengyi 1 Jiu , a,c Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. zdb rrtsadanra ie ucin uha lvtdlvrezmsadicesdtesrmtotal serum the increased and enzymes liver elevated as character- such which function, disease, liver liver abnormal pregnancy-specific reversible and a pruritus is by (ICP) ized pregnancy of The cholestasis manuscript. Intrahepatic the writing and analysis data collection, Introduction samples funders. design, of study independent fund- the was 20202BABL216010 The in team (No. role research (No.2018M642594). Province no Grant Jiangxi Foundation had of Science body Foundation Postdoctoral ing Science China National and the No.20192BBG70003) by and supported was study This Funding disease. pregnancy of cholestasis abstract Tweetable patients. ICP for expression treatment HDL gene new and provide and TG disease, Keywords CHOL, ICP controls. DBIL, of local diagnose AST, and genetic two ICP TBA, to with in contribute of mutation patients no value the mutation, average Moreover, one Conclusion have higher which women. of have patients pregnant level genes the expression healthy to family the than compared tissue, ABC higher placental in significantly in was And mutations patients chains. side ICP acid in amino binding ATP-ligand functional of known the in located which reference, identified were mutations including pathogenic possible genes novel 7 which, of results, Results measures outcome Main MAF frequencies: allele Methods Samples Setting approach. (WES) Design sequencing whole-exome by disease (ICP) pregnancy Objectives [email protected] Zou: Yang [email protected] Zheng: Jiusheng [email protected] Wu: Meiling [email protected] Liang: Zhenyi [email protected] Nie: Liju [email protected] Zeng: Xiaomin [email protected] Li: Zengming [email protected] Xin: Siming [email protected] Lai: Hua [email protected] Liu: Xianxian E h N,adcnutascainbtengntcvrat n oa ieacids. bile total and variants genetic between association conduct and DNA, the WES Jiangxi. ABCC2 edtce 2wr oe.W lsie hs oia orpnl codn oteprediction the to according panels four as loci these classified We novel. were 42 detected We 5 C patients. 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Data may be preliminary. obntra rb nhrLgto cA)mto.Ec eutn ulfidcpue irr a then using was constructed library were captured library qualified DNA resulting platforms. Each exome sequencing protocols. out BGISEQ-500 carry method. manufacturer’s to on (cPAL) to loaded subjected Ligation according were Kit Anchor samples the Kit prep Probe patients of Exon ICP Mini combinatorial USA) integrality 151 BGI Fisher, DNA and of (Thermo with Genomic concentration total spectrophotometer A the capture Blood Nanodrop-1000 respectively. genotyping, by Prep electrophoresis, determined for gel were Axy requirements and samples by the DNA exacted blood fulfill the To peripheral all 05119KC3). from exacted No. was (Item DNA controls. genomic were negative 151 amount), as bleeding ICP and without sequencing weight women (birth exome pregnancy newborns Whole 1029 distribution and recruited cell lipoprotein: women also low-density blood pregnant DBIL, we HDL, red of Besides, bilirubin: PLT, lipoprotein: outcomes direct recorded. high-density blood platelet: and TBIL, TG, routine RBC, acid) (CHOL, bilirubin: P), uric index cell: total Mg, LDL, lipid AST, blood Ca, IDBIL), ALT, red Cl, (TBA, 26 bilirubin: WBC, Na, index indirect addition, (K, pruritus cell: function concentration In features liver skin blood ion RDW.SD), basic white of BMI), etc. patient width.SD: of including basis AST, index: counts data, mass and the (the neonatal ALT body on and test and TBA, maternal ICP age of e.g. as gestational indicators (age, main indexes, diagnosed six biochemistry disease, covering liver features liver clinical abnormal other with excluded combination were in women pregnancy 151 data clinical outcomes. and patients pregnancy Patients 151 and in for data transport clinical especially acids the bile mutations, to in of Methods involved them genes presence related series the and ABC disease extensively of ICP investigate variants, transporter with functional to acids susceptibility new WES bile confers of transporters of discovery performed ABC the genes we in variation series Therefore, genetic ABC ICP. the the that to have hypothesized in there we variants background, knowledge, genetic this best Given the our identify to disease new to now, ICP for disease. 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(ABC) ABCG. and cassette ABCF for ATP-binding ABCE, Except of ABCD, ABCC, function ABCB, physiological ABCA, ABCB4 key ICP. implicating members: of the ICP, distinct pathogenesis seven with is covered the patients transport in of role salts pivotal sisters Bile a and playing ICP mothers salts condition. to in bile the incidence of lead for transporters high may predisposition a genetic acids showed a backgrounds. studies, that bile environmental and pedigree of genetic in excretion hormonal, ysis including and factors, multiple secretion on important. transport, very metabolism is stillbirth. disease. complications synthesis, and fetal the abnormal morbidity with fetal The associated of is fetal risk and and the ICP distress increased of fetal mechanisms TBA scores, of Apgar level low the distress, respiratory birth, differences. death. preterm geographical spontaneous on including period. depending comes, postpartum 15.6% early and the 0.1% in delivery after 10 completely ([?] (TBA) acids bile 3-7 13 , ti oe htapoiaey2-%o C rgace r ffce yftlmortality. fetal by affected are pregnancies ICP of 2%-4% approximately that noted is It ABCB11 hrfr,teeilg otiuigt h eeomn fIPdsaei ope htdepend that complex is disease ICP of development the to contributing etiology the Therefore, ABCB4 and , ABCB11 ABCC2 μ o/) perdi h eodadtidtietro h rgac n resolved and pregnancy the of trimester third and second the in appeared mol/L), and r ucinlkongnshvn neeto h eeomn fICP. of development the on effect an having genes known functional are ABCC2 ee,terl fohrACtasotr’gnssest less to seems genes transporters’ ABC other of role the genes, 15-18 mn hc,tegn uain ntehepatocellular the in mutations gene the which, Among 3 ,2 1, 1 h niec fIPdsaerpre between reported disease ICP of incidence The C a soitdwt h des ea out- fetal adverse the with associated has ICP 10-12 hrfr,utnln h genetic the untangling Therefore, 19 14 aiilcutrn anal- clustering Familial ,9 8, And Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. tpgie satls,3 ’pie T,2 ’pie T,1 ontemgn,3 ptemgn,3 gene, upstream 32 gene, downstream 10 13 UTR, splice, primer 76 5’ synonymous, 21 297 UTR, intron, primer 2057 3’ covered 36 variants of lost, types /start These gained 12 in stop genes. mutations series 1254 of ABCD-ABCG including cohort in variants, a genetic in 2953 variants identified of identify - we consisted to Overall, which WES genes targeted disease. 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ABCC8 - ABCC12 ABCC2 ABCB1 cor.test 0 variants 408 , Ser1342Tyr) , function ABCB4 Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. 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Chimera UCSF using simultaneously compared were gene ABCA5 l46h and Ala456Thr 0 21 20, Cys2440Tyr, Asn2492Ser, e42VladTr92l) l36y n l39a nall in Ile339Val and Glu386Lys Thr4912Ala), and Leu4624Val ABCC1 hsgn a w anncetd idn oan which domains binding nucleotide main two has gene This Val997Ala, ABCC12 ABCC2 5 Ser497Gly, ABCA13 ABCG2 ABCA13 ABCB11 e14Trmutation Ser1342Tyr Pro37Ser), ABCA7 < .1i l aaae) 2oto 2 variants 320 of out 42 databases), all in 0.01 e66e.Tefeunyo hs muta- these of frequency The Leu646Met. ABCC3 e31Anand Ser3111Asn l19Tr l65r n Leu589Met, and Gln605Pro Gln1194Ter, Ser3286Ter, Pro449His, ABCD4 Leu137Phe, e35h n Thr385Ala, and Ser395Phe ABCB1 ABCA8 P ABCG1 1.10E-14). = ABCC5 Pro693Ser, Val8Ala, h38l.Only Thr378Ile. Gln851Pro, ABCA10 ABCA3 ABCB5 P = Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. usqetfntoa eicto.Fnly obn E n lncldt,i si ao fdcpeigof deciphering of favor in is and it data data, clinical rel- clinical and disease. are scale and mutations ICP patients large WES between of these relatively combine mechanism analysis of the Finally, a molecular data association unraveled from clinical verification. for have women the functional support studies Moreover, pregnancy subsequent data no China. among providing date, in disease complete, to (n=151) WES- hepatic atively Second, first sample of representative the disease. genes of nationally gynecological mutations of ICP ABC genetic and with in the investigated mutations associated obstetric results genetic genes Our in family genes. spectrum ABC series identified ABC MAF proved other the in has variations which across novel technology, variation WES genes of sequence advent functional the unearth following to first, diseases. efficient strengths: major be 4 to 44 has covering study loci present pathogenic new The novel 42 disease. identified potential ICP we uncover with all, diagnosed to In andlimitations were Strengths technology transport. which acid patients sequencing bile 151 whole-exome in in involved use genes genes ABC to family first ABC in the mutations is study This findings Main the ICP, healthy of and Discussion characteristic genes transporters’ clinical mutations ABC two a of with local mutations as and controls. with ICP TBA, ICP samples healthy ranked: particular, in ICP AST, were measured in In which controls TBA, mutation of no local higher 3). value 113 average (Figure exhibited mutation, the mutations population one of with trend two healthy patients 414 both 31 in contained than HDL controls patients and ICP TG that six ICP abnormal CHOL, confirms the DBIL, that with which with value, found patients associated reference we 151 acids the Additionally, bile of elevated than and TG S2. 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Figure material, Supplementary (See tissue placental in changed have , sn hsmto,w ucsflyietfidnvlcniaeptoeiiylc ihknown with loci pathogenicity candidate novel identified successfully we method, this Using ABCB4 , ABCB11 and ABCC2 ABCC2 eaaye h RAepeso ee of level expression mRNA the analyzed we , nadto oteetregns eas i some dig also we genes, three these to addition In . > 6 C ihoemutation one with ICP P < > .5 ieec ntegene the in difference 0.05) C ihn mutation no with ICP ABCC2 ABCC6 ABCC2 upregulated P , < ABCE1 eein gene 0.05). > Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. 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Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. X Z N LadM olce ape.LM ZadY otiue otecnetaddsg fthe of design and concept the to contributed YZ and manuscript. the JZ drafted LZM, and samples. figures collected the MW prepared and data, ZL the LN, analyzed XZ, experiments, the SX, performed HL and XL exist. interest of authorship conflicts to potential Contribution no that clinical declared have for authors variant The candidate potential suggest new interests provide and of findings disease are Disclosure Our ICP that of variants investigated. further ( basis genetic TG be genetic of and diagnose. to the CHOL detection needs into DBIL, results the validation insights AST, The ALT, functional found valuable TBA, their we sites. including Nevertheless, pathogenic Moreover, index, them, novel biochemical 0.05). these conservative. six Among with of were associated some genes. loci significantly on family most analysis ABC that bioinformatics altogether showed and 44 validation in performed mutations We in identified pathogenic were potential loci novel seven 42 in detected which, We diagnosed, evidence disease. the genetic ICP provides timely study the Our to to Conclusion contribution person. mutations susceptible potential these ICP have for mutations genotyped treatment these to and that attention mutations important immediate and get is to concentrations it order salt outcomes, bile fetal in and variation injury. to liver whether potential drug-induced in of has interest ICP pathogenesis much of to ICP currently statue lead in is genes involved there transporters’ mutations Recently, ABC genetic patients. in ICP the mutations as for on the (such therapies work novel disease. facts genes inspire further ICP the functional for Therefore, known support studies the results disease. analysis Our of genetic ICP. basis reports with molecular of associated the handful enrich a only only ABCB11 not absorption now findings metabolism. are lipid present lipid there to The and date, essential disease to is ICP surprisingly, nature, on Not genes amphipathic family its ABC for the known researchers in result, human. sug- well mutations this in which which with balance Consistent index, (BA), energy abnormality. biochemical acids and lipid was to bile type lead ICP that, mutant could in TBA and found results of type accumulation level is the wild TBA situation that between of gested this difference accumulation of significant this explanation the and reasonable Moreover, level, controls. The TBA healthy HDL. on in and effect mutation TG additive no CHOL, exacerbation. has and DBIL, mutation genes AST, double transporter the that ABC did in level so mutation TBA expected, average no As the mutation, increased one mutations two mutations, 50.32 genotypes’ two both and of harbored 20.22 analysis ICP summation 17.33, Notably, the a by that once. having genes at suggested of mutations al disease. two polymorphism ICP TG et contain of of variants CHOL, K development mutated DBIL, the Piatek the on AST, Furthermore, of effect TBA, possibility ICP 3). the higher among to Figure with identified pointed and associated co-existence were S2; was activity (Table variants AST novel HDL and 42 and ALT containing TBA, group genes. serum mutation transporter ABC in the in genes. elevations mutations family the have ABC who that patients other suggested many al with understanding et the along Bacq strengthens disease, and ICP mutations of of role basis the genetic extended findings of These group. damaging especially ABCB4 and ABCC2 / ABCB11 u loepn h e addt ee AC,AC,AC,ABCD-ABCG) ABCC, ABCB, (ABCA, genes candidate new the expand also but ) μ 36 o/,rsetvl,bsdo h ieec naeaeTAlvl nIPwith ICP in levels TBA average in difference the on based respectively, mol/L, ABCB4 / 33-35 utemr,cnieigtengtv ffc fteACmttoso mother on mutations ABC the of effect negative the considering Furthermore, ABCC2 hs ae oehr u td rvdsadtoa upr o ffc of effect for support additional provides study our together, taken Thus, , ABCB11 ohrnwcniaegnscudb imre o aiu forms various for biomarker a be could genes candidate new /other 32 ossetwt hsrsl,i u td,w aesxIPpatients ICP six have we study, our in result, this with Consistent and ABCC2 8 31 nareetwt ht u eut ofimdthat confirmed results our that, with agreement In n h eann 4lc eei te genes. other in were loci 34 remaining the and , ABCB4 P < , Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 6 ozahR,SvkD,BanW.Fmla eurn nrhptccoetsso rgac:agenetic a trait. dominant pregnancy: sex-limited, of a cholestasis of intrahepatic transmission recurrent Familial for WE. evidence Braun providing DA, study kindred. Sivak one RT, in Holzbach pregnancy 16. of FXR cholestasis intrahepatic sensor of Spectrum. acid Inheritance Evolving S. bile An 1993;43:315-7. Kivinen central Disease: ML, Liver Hirvioja the Human 15. of in variants Aberrations Functional Gene Dis ABCB4 Liver F. al. Semin Lammert et MC, PH, Reichert Dixon 14. pregnancy. A, maternal of cholestasis Milona pregnancy: intrahepatic SW, of in identified Mil cholestasis levels. Van Intrahepatic acid intrahepatic al. bile in 13. elevated et asphyxia with GC, fetal associated Page-Christiaens Predicting outcomes MP, S. fetal Koster Tinar and L, R, Brouwers Borekci S, 12. Okcu O, case-control Oztekin pregnancy. I, population-based of Aydal cholestasis prospective D, intrahepatic a Oztekin severe of 11. outcomes: Association C. pregnancy Williamson adverse M, Knight with PJ, study. Steer pregnancy pregnancy: PT, of of Seed LC, cholestasis cholestasis Chappell Intrahepatic V, TM. Geenes Goodwin 10. management. M, expectant Ames-Castro with associated JG, outcome Ouzounian cholestasis.perinatal OM, obstetric compli- Alsulyman in pregnancies outcome in 9. Fetal outcomes cohort. GN. for Fetal California Storey management L. Northern NM, aggressive Bull a Fisk P, of in 8. Rosenthal pregnancy era P, of Bacchetti an cholestasis A, during intrahepatic Caughey outcomes by J, cated Vargas Pregnancy M, al. Rook et 7. S, Ingles pregnancy. KM, of pregnancy cholestasis Kwok of intrahepatic cholestasis RH, intrahepatic Lee of prevalence population. 6. The Angeles M. Los Incerpi Latina J, primarily Greenspoon a outcome TM, in cholestasis, Goodwin Obstetric RH, cases. AH. Lee 70 Shennan 5. RM, of Tribe series C, a Williamson management: J, patho- active Girling molecular with CN, Piercy pregnancy: Chile. AP, of Kenyon in cholestasis 4. pregnancy Intrahepatic S. of Matern cholestasis management. A, and intrahepatic Glantz diagnosis of HU, genesis, Prevalence Marschall F, expectant Lammert al. of et 3. week J, additional Ribalta each Med MC, by Intern Gonzalez death Ann fetal H, and Reyes infant of 2. risk The age. gestational al. by et pregnancy e1-5. of J, cholestasis Page intrahepatic E, in Kim management A, Puljic 1. each by and approved China, was in approval Hospital References ethics Health the Child consent. and and informed Maternal Declaration gave Provincial woman Helsinki Jiangxi participated the of Board of Review tenets Institutional the the authors followed All study present manuscript. The the revised approval and ethical data of the Details manuscript. analyzed final experiments, the the approval performed and YZ read and JZ study. Hepatology 1978;88:487-93. 2018;38:299-307. 2014;59:1482-91. rhGnclObstet Gynecol Arch Hepatol J mJPerinatol J Am BJOG 2009;280:975-9. Perinatol J 2000;33:1012-21. Gastroenterology 9 2002;109:282-8. 2008;25:341-5. 2006;26:527-32. mJOse Gynecol Obstet J Am mJOse Gynecol Obstet J Am rJOse Gynaecol Obstet J Br 2007;133:507-16. mJOse Gynecol Obstet J Am Gastroenterology LSOne PLoS 1996;175:957-60. 052210e1-7. 2015;212:100 1988;95:1137-43. 2012;7:e28343. 1983;85:175-9. 2015;212:667 lnGenet Clin Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 5 agD,Tzm ,ChnD,CryM.Feigntrlhdohlcbl cd niisintestinal inhibits acids bile hydrophilic biological natural to mouse. Feeding gallstone-susceptible relationship MC. the Carey their in DE, and studies Cohen acids absorption: S, cholesterol Tazuma bile DQ, of and Wang mechanisms properties 35. problem. - Physicochemical the signalling of A. overview insulin Roda an and AF, properties: metabolism Hofmann glucose in the 34. in acids polymorphism Bile gene RA. transporters’ Haeusler needs. ABC research TR, of Ahmad role of The cholestasis 33. al. pregnancy. intrahepatic et of treatment. cholestasis in J, to intrahepatic Magielda therapy of G, response acid etiology Kurzawinska of Ursodeoxycholic K, predictive Piatek al. factors 32. et and AI, conditions Lecuyer real-world M, Dis in Besco pregnancy. Results le of Y, pregnancy: cholestasis Bacq during progesterone trio 31. altered the mother-placenta-foetus on treatment the acid in ursodeoxycholic of homeostasis Effect Pharmacol nuclear acid al. the et preventing bile L, Rivas by and MJ, chemosensitivity Monte regulates MC, Estiu MicroRNA-31-5p 30. al. carcinoma. et hepatocellular transporter Y, in drug PARP1 You and of Y, receptor location Zhou nuclear KT, alters Que cirrhosis Alcohol 29. susceptibility al. to et liver. ABCB11 AC, human of Donepudi in alleles Q, variant expression Cheng of VR, Contribution More al. 28. et pregnancy. J, of Chambers cholestasis SW, intrahepatic Mil to recur- van causing PH, mutation Dixon gene 27. ABCB4 pregnancy. of heterozygous of Novel cholestasis cholestasis MP. intrahepatic intrahepatic Nageotte severe first-trimester ML, of rent Stephenson management RC, Successful Johnston case. al. Japanese 26. first et a N, of Kamimura report H, Abe pregnancy: K, Kamimura 25. pregnancy. Hydrocephalus of Fetal cholestasis Causing intrahepatic of Gene pathophysiology L1CAM The the Gastroenterol C. in Williamson Mutation PH, Silent Dixon Novel 24. A al. et Sequencing. genetic M, Whole-Exome a Chen by as Y, Detected HSPA1L Li reveals Y, sequencing Sun exome 23. Whole birth. preterm al. et spontaneous BE, for Graham factor MK, risk Karjalainen JM, and Huusko multispecific specificity activity. the substrate 22. transport in alters Trp1254 methotrexate (ABCC2), of of (MRP2) Mutation SP. loss 2 Cole in protein RG, rat resistance results Deeley multidrug with MZ, Vasa transporter, BSEP C, anion Westlake human organic glycine- CJ, Oleschuk of of K, vesicles comparison Ito by a 21. Transport Y. 3-sulfate: Sugiyama taurolithocholate AF, Hofmann and R, salts Bsep. Onuki cholestasis bile intrahepatic H, taurine-conjugated in Suzuki heterogeneity T, and index Takada of H, of evidence Hayashi Genetic sisters 20. in al. et cholestasis K, obstetric Avela pregnancy. of A, of Risk Ropponen M, S. Savander kindred. Saarikoski 19. large T, a Mononen in S, pregnancy Heinonen patients. of ML, cholestasis Eloranta Idiopathic 18. M. Gonzalez-Ceron J, Ribalta 1976;17:709-13. H, Reyes 17. 2017;49:63-69. ici ipy Acta Biophys Biochim lnGenet Clin 2015;79:316-29. 2016;40:141-53. Gut a e Endocrinol Rev Nat 2003;52:1025-9. 2001;60:42-5. rgMtbDispos Metab Drug 2005;1738:54-62. 2019;15:701-12. rn Genet Front Gut ii Res Lipid J M Gastroenterol BMC LSGenet PLoS 2009;58:537-44. 2013;41:1148-55. x lnCne Res Cancer Clin Exp J ieo Pol Ginekol ilChem Biol J 1984;25:1477-89. 2019;10:817. 10 2018;14:e1007394. Perinatol J 2018;89:393-97. 2014;14:160. 2001;276:38108-14. mJPyilGsrits ie Physiol Liver Gastrointest Physiol J Am 2014;34:711-2. 2018;37:268. lnRsHepatol Res Clin rJClin J Br i Liver Dig Gut Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 42 41 40 39 38 37 36 35 34 33 32 31 30 29 28 27 26 25 24 23 22 21 20 19 18 17 16 15 14 13 12 SIFT 11 change Protein 10 Alleles 9 8 Position 7 Chr 6 5 Patients 4 3 2 1 Gene ICP with Chinese ID Han 151 in identified were mutations novel ABC disease. pathogenic potential Forty-two Protein Resistance 1. Multidrug Table Pharmaceuticals. of 125 Role Using the Injury Evaluating Liver AD. Drug-Induced Rodrigues 2017;30:1219-29. Predicting PD, in Bonin K, Inhibition He (MDR3) F, 3 Shah MD, Aleo 36. 2003;285:G494-502. ABCA13 ABCG1 ABCA13 ABCA12 ABCA10 ABCA3 ABCG1 ABCF2 ABCD4 ABCD4 ABCC12 ABCC9 ABCC6 ABCC5 ABCC3 ABCC1 ABCB9 ABCB9 ABCA13 ABCA13 ABCA12 ABCA10 ABCA8 ABCA7 ABCA5 ABCA2 ABCA2 ABCG2 ABCC9 ABCC3 ABCC2 ABCB11 ABCB11 ABCB11 ABCB5 ABCB4 ABCB4 ABCB4 ABCB1 ABCA13 ABCA12 ABCA4 a,b a,b a,b a,b a,b a,b,c a,b a,b a,b a,b a,b a a,b C5 h74386 / l26Vl096()1()06 T eta .200(*09 .6 111 .3Asn n10GEA/bN/hnMPdatabases 1000G/ExAC/dbSNP/ChinaMAP in Absent 0.13 databases 1000G/ExAC/dbSNP/ChinaMAP in (1/151) Absent 0.66% 0/(2*1029) 3.74E-08 (5/151) 3.31% 0/(2*1029) 0 0 4.92 0.011 0.354 0.005 0.999 4.17 1 databases 1000G/ExAC/dbSNP/ChinaMAP in 1 Absent 2.92 5.59 6.17 5.79 1 0.875 4.22 0.974 Neutral 4.81 1 damaging Pos 1 1 (T) 2.20E-16 1 0.66 (D) 4.84 -1.98 4.84 (21/151) 13.91% 5.45 damaging 1 Pos damaging damaging Pos Pos 1 damaging 4.95 0.979 Pos (D) -2.19 (D) damaging -2.11 5.38 5.52 0/(2*1029) Pro (D) 5.62 -3.3 (D) -2.28 1 (N) 1 (N) (T) 1 databases damaging 0.52 1000G/ExAC/dbSNP/ChinaMAP Pro 0.702 (T) in 5.32 0.916 Absent 1 (T) 5.43 0.21 (D) (N) -2.77 0.018 damaging 0.999 0 Pro damaging (N) Pro (N) 0.958 5 (T) Ile2460Val 1 0.071 damaging (D) Pro (N) -2.63 (T) 1 5.7 (D) 0.403 Thr378Ile -2.77 (T) A/G damaging 0.09 5.35 (T) (D) 1 Pro 0.126 -2.76 (D) Ser3111Asn 1 1 5.73 0.94 (D) damaging damaging 1 0.999 Pro Pro C/T 48318169 damaging 0.993 Asn2492Ser Pro 1 (D) (D) 4.84 1.10E-14 -2.61 1 0.002 3.05 (T) G/A 0.156 Leu823Val (D) chr7 43708158 (D) -3.35 (T) (16/151) 0.990 Ser593Gly 0.85 T/C 10.60% (T) (N) damaging 0.95 0.983 chr21 48349554 Pro (T) 1 0.959 (D) 215802301 G/C 0.254 damaging 0.812 1 (D) Pro T/C 0.033 5.45 Ile273Val chr7 (D) -2.71 5.55 Gly47Ser ICP52 (T) 4.31 chr2 67181648 (D) 0.217 damaging 0.999 Pro 5.7 5.72 0/(2*1029) 5.29 ICP22 (D) chr17 A/G 2348506 (N) 0.835 -2.6 Thr385Ala 0.999 damaging (T) 4.2 C/T Pro 0.999 0.111 (D) ICP111 Ser395Phe chr16 0.999 damaging (D) (D) 43704752 Pro 150923406 0.029 -2.4 0.999 (D) ICP71 damaging T/C (D) (D) 1 Pro Pro37Ser 0.026 6.11 -5.88 chr21 (D) G/A (D) 5.79 5.2 Glu1304Val 0.972 -2.477 ICP64 chr7 damaging 0.999 (D) damaging Pro 0.011 74757168 Pro (D) His1043Gln ICP22 (T) -3.77 G/A 74757137 (D) 0.268 0.999 chr14 1 T/A (D) Gln851Pro ICP113 (D) chr14 -2.41 -4.44 5.5 (T) 48180227 3.91 (D) G/T 0.176 0.682 Leu137Phe ICP84 21968809 (N) damaging chr16 0.958 Pro 5.61 T/G (N) damaging (T) ICP142 Ser497Gly chr12 16259657 0.954 Pro 0.891 (D) C/T 183670989 0.001 (D) 5.47 ICP95 (D) chr16 (D) 0.994 Damaging -1.77 0.017 damaging (D) A/G 1 Ile339Val Pro -2.17 (N) ICP155 Damaging Damaging chr3 48734467 Damaging (T) 0.984 5.91 0.093 (D) Glu386Lys -2.33 ICP36 Thr4912Ala chr17 16149964 (D) (D) (D) 0.009 T/C -3.27 Leu4624Val (D) (N) -2.57 -2.58 1 ICP57 chr16 C/T 123433209 A/G Damaging (D) ICP49 Ile1022Thr (D) Damaging 0.015 Damaging C/G 0.961 chr12 123430667 (D) ICP57 0.999 48634399 (T) chr12 - 5.66 (D) (D) A/G 48559709 0.527 (D) -2.95 -2.82 ICP98 Val8Ala (T) 0.998 0.351 (N) chr7 0.999 215865543 Damaging Damaging Pro449His ICP140 chr7 (D) (D) A/G 0.01 0.043 ICP112 Damaging (D) chr2 (D) - (D) 1 Val997Ala -1.99 1 C/A (D) ICP107 Ala583Val -2.5 66938153 Damaging (D) (D) -2.35 (D) 0.001 (D) Asp1108Glu ICP18 0.999 chr17 0.994 A/G Leu646Met (D) 0.0 1045131 G/A (D) Damaging (A) ICP95 -2.65 1 chr19 67266794 G/C 139913419 (D) G/T (D) 0.0 (D) 0.0 (D) Ala456Thr -2.21 chr17 0.972 139910497 (D) Ile1147Thr ICP47 chr9 0.786 89013418 (A) (D) 1 0.006 ICP108 chr9 C/T (D) Ser1342Tyr 0.002 Leu589Met chr4 T/C Damaging ICP102 (D) - 1 22061100 ICP19 48755166 C/A G/T (D) (D) Gln605Pro chr12 -4.24 1 ICP64 chr17 101605418 (D) 169826599 0.0 Gln1194Ter Ser925Ile (D) (D) 0.002 ICP6 1 - chr10 T/G chr2 ICP124 (D) G/A 169826057 G/T 0.031 Gly527Glu ICP93 169783704 Lys386Glu (D) chr2 Trp708Ter 0.999 20767985 ICP79 chr2 (D) C/T 0.001 ICP2 Pro693Ser chr7 T/C C/T ICP118 87069134 87073053 G/A 87053310 ICP115 chr7 Phe754Ser ICP47 chr7 chr7 87173579 ICP133,135 A/G chr7 ICP154 94522278 ICP21 ICP121 chr1 ICP76 C3 h1 7105AGLu6Sr008()1()-.8()Podmgn .10.003 3.21 1 damaging Pro (D) -2.28 5.68 1 (N) 1 (D) 0.038 5.49 Leu260Ser Damaging A/G 67212035 chr17 - ICP35 Damaging (D) -3.67 (A) 1 (D) 1 - (D) 0.041 Ser3286Ter Cys2440Tyr C/G C/T 48354004 215809749 chr7 chr2 ICP112 ICP75 11 d hmRsToxicol Res Chem uainTaster Mutation e A HMCmrhniepeito result prediction Comprehensive FATHMM f GERP++NR g PhastCons100way ver h A ncnrl A n11ICP 151 in MAF controls in MAF P au cnrl-C)MFi databases in MAF (controls-ICP) value Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. ereaddrcino orlto coefficients. correlation of direction and degree < tissue. S5. placental Figure in patients ICP 4 and healthy S4. Figure species. vertebrate in conserved highly were genes. S3. ABCD-ABCG Figure (D) Total and genes; before QCSNPs: ABCC variants (C) control; genetic genes; quality ABCB of before number SNPs total S2. Total SNPs. The Figure Novel TSNPs: (B) NSNPs: variants. control; genes. novel quality series after the ABC SNPs and in control, variants quality genetic after of types the of S1. Figure S2. Table of end the at section S1. Information Table Supporting the in online found article. loci. be the may the conservative information more supporting the Additional values, the higher the information and Supporting 1, to 0 from ranged score h g f e d c b a r aaig rbbedmgn;Psdmgn:Psil damaging. Possible damaging: Pos damaging; probable damaging: Pro :plmrhs,A ies asn automatic. causing disease A: structure. polymorphism, protein N: encoded the on mutation this of change the of effect The EP+R RP+cnevto cr.Tehge h au,temr osraieteloci. the conservative more the value, the higher The score. conservation GREP++ GERP++NR: sequencing. Sanger by validated was loci This PhastCons100way :dsaecuig :tolerated. T: disease-causing, D: analysis. conservative evolutionary by performed was loci This .5 orlto offiinsbtenec aro etrs t ieadclr eaaeyrpeetthe represent separately colors and size Its features. of pair each between coefficients correlation 0.05) ecitv ttsiso 6ciia aao C niiul soitdwt hs mutations. these with associated individuals ICP of data clinical genes. 26 transporters’ of ABC statistics in Descriptive variants the confirmed for used Primes h vltoaycnevtv nlsso B oe ersnaiemttos hr mutations There mutations. representative novel ABC of analysis conservative evolutionary The orlto offiinsaogtet-i lnclfaue.Tedt niaetesgicn ( significant the indicate dots The features. clinical twenty-six among coefficients Correlation h itiuinadnmeso eei ainsfo E aafrIP A h percentage The (A) ICP. for data WES from variants genetic of numbers and distribution The agrsqecn ovldt h oe ainsi B eisgns A BAgns (B) genes; ABCA (A) genes. series ABC in variants novel the validate to sequencing Sanger h oprsno h xrsinlvlof level expression the of comparison The e:cnevtv rdcini etbae n10vrerts h crso PhastCons of scores The vertebrates. 100 in vertebrates in prediction conservative ver: 12 ABCC6 , ABCE1 and ABCG5 ee ewe two between genes P Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 13 Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 14 Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741603.33274306 — This a preprint and has not been peer reviewed. Data may be preliminary. 15