Tshawb nrhiav MUC1 Variants hauv Cov Neeg Mob Clinicopathologically Diagnosed Nrog Autosomal Dominant Tubulointerstitial raum Kab Mob

Taw qhia

Autosomal dominant tubulointerstitial raum kab mob (ADTKD)-MUC1 feem ntau yog tshwm sim los ntawm kev hloov pauv hloov vim yog ib qho kev nkag mus rau hauv qhov sib txawv ntawm tus lej tandem rov ua dua (VNTR) cheeb tsam hauv MUC1. Vim yog qhov nyuaj ntawm qhov sib txawv ntawm hotspot, kev txheeb xyuas siv cov ntawv nyeem luv luv (SRSs) yog qhov nyuaj. Txawm hais tias cov kev tshawb fawb tsis ntev los no tau qhia txog qhov muaj txiaj ntsig ntawm cov ntawv nyeem ntev ntev (LRSs), qhov sib txawv ntawm MUC1 ntau yam hauv cov neeg mob uas xav tias ADTKD tseem tsis paub. Peb tsom kom paub meej qhov kev nthuav dav no thiab cov yam ntxwv ntawm caj ces thiab kev kho mob tshwm sim ntawm ADTKD-MUC1 hauv cov neeg Nyij Pooj siv SRS thiab LRS.

Cov txheej txheem

Txij Lub Ib Hlis 2015 txog Lub Kaum Ob Hlis 2019, kev tshuaj ntsuam caj ces tau ua los ntawm kev siv SRS hauv 48 tus neeg mob uas kuaj mob ADTKD. Kev tshuaj xyuas ntxiv tau ua los ntawm kev siv LRS hauv cov neeg mob uas tau txais SRS tsis zoo.

Cov txiaj ntsig

Cov qhabnias luv luv nyeem tau qhia MUC1 qhov sib txawv hauv 1 tus neeg mob harboring ib qho cytosine ntxig rau hauv chav tsev thib ob ntawm thaj tsam VNTR; Txawm li cas los xij, cov cheeb tsam VNTR tob tuaj yeem tsis tau nyeem los ntawm SRS. Yog li ntawd, peb tau ua qhov kev soj ntsuam ntev ntev ntawm 39 tus neeg mob thiab kuaj pom MUC1 VNTR sib txawv hauv 8 tus neeg mob (tag nrho, 9 tus neeg mob los ntawm cov tsev neeg tsis sib xws). Nrog rau kev suav nrog cov neeg mob hauv tsev neeg (n ¼ 31), lub hnub nyoog nruab nrab ntawm kev loj hlob ntawm cov kab mob raum kawg (ESKD) yog 45 xyoo (95 feem pua ​​​​CI: 40-40 xyoo).

Xaus

Hauv Nyij Pooj, qhov kev kuaj pom tus nqi ntawm MUC1 qhov sib txawv hauv cov neeg mob uas tau kuaj pom ADTKD yog 18.8 feem pua. Ntau tshaj 20 feem pua ​​​​ntawm cov neeg mob uas tsis zoo SRS cov txiaj ntsig tau muaj MUC1 qhov txawv txav los ntawm LRS.

Ntsiab lus

ADTKD; ADTKD-MUC1; nyeem ntawv ntev; MCKD; NGS; SMRT sib txuas.

Nyem qhov no kom tau txaisCov txiaj ntsig Cistanche

ADTKD yog ib pawg ntawm cov kab mob hauv lub raum uas tshwm sim los ntawm kev loj hlob ntawm tubulointerstitial fibrosis thiab tubular atrophy ua rau ESKD. ADTKD tuaj yeem muab faib ua cov subtypes raws li qhov ua rau muaj kev cuam tshuam ntawm caj ces, nrog rau qhov txawv txav hauv UMOD, MUC1, REN, HNF1B, thiab SEC61A1. Muab qhov tsis tseem ceeb ntawm cov yam ntxwv sib koom ua ke ntawm ADTKD, suav nrog cov urinary sediment txawv txav thiab me me rau qhov tsis zoo proteinuria, ADTKD tau suav tias yog qhov tsis txaus ntseeg.

MUC1 yog ib qho transmembrane glycoprotein tau nthuav tawm nyob rau hauv apical membrane ntawm tuab ascending limb ntawm lub voj ntawm Henle, distal convoluted tubules, thiab sau ducts. Feem ntau cov caj ces ntawm ADTKD-MUC1 yog qhov hloov pauv hloov pauv los ntawm kev ntxig ib lub hauv paus rau hauv VNTR polymorphism ntawm MUC1 (OMIM 158340; 1q22) hauv exon 2.5 Qhov kev hloov pauv no txiav tawm cov khoom sib xyaw ntawm MUC1 protein tom qab thaj tsam VNTR thiab tsim cov protein luv luv (MUC1fs) uas tsis muaj C-terminal domain (Daim duab 1a). MUC1fs ces accumulates nyob rau hauv lub cytoplasm vim qhov txawv txav intracellular protein thauj. Lub cheeb tsam VNTR nthuav tawm ntau qhov sib txawv ntawm tus kheej vim tias nws yog tsim los ntawm 60-nucleotide units uas rov ua dua 20 txog 125 zaug, nrog rau txhua tus allele muaj qhov tshwj xeeb rov ua dua (Daim duab 1b). Yog li ntawd, nrhiav cov noob variants siv SRSs tau nyuaj vim qhov nyuaj ntawm qhov sib txawv ntawm qhov hotspot.

Cov kev tshawb fawb yav dhau los tau siv lwm txoj hauv kev, suav nrog huab hwm coj spectrometry thiab kev sojntsuam txuas ntxiv raws li tau hais, txhawm rau txheeb xyuas qhov sib txawv ntawm qhov sib txawv; Txawm li cas los xij, cov txheej txheem no yog technically xav tau. Cov kev tshawb fawb tsis ntev los no tau pom qhov muaj txiaj ntsig ntawm LRSs, uas siv tib lub sijhawm ntawm lub sijhawm tiag tiag (SMRT) kev sib txuas los txheeb xyuas MUC1 VNTR sib txawv. Txawm li cas los xij, MUC1 VNTR variants tseem tsis tau muaj tus yam ntxwv ntawm cov neeg Nyij Pooj. Tsis tas li ntawd, qhov sib txawv ntawm MUC1 qhov sib txawv hauv cov neeg mob uas xav tias ADTKD tseem tsis tau paub. Yog li ntawd, lub hom phiaj ntawm txoj kev tshawb no yog los qhia meej qhov kev nthuav dav no, suav nrog cov yam ntxwv ntawm caj ces thiab kev kho mob, ntawm ADTKD-MUC1 hauv cov neeg Nyij Pooj. Ntawm no, peb kuaj tau 9 tus neeg mob nrog ADTKD-MUC1 uas muaj VNTR sib txawv kuaj pom siv SRS (n ¼ 1) lossis LRS (n ¼ 8) ntawm 48 qhov chaw kho mob xav tias ADTKD.

Cov txheej txheem

1. Cov neeg koom nrog

Nyob nruab nrab ntawm Lub Ib Hlis 2015 txog Lub Kaum Ob Hlis 2019, peb tau soj ntsuam cov noob caj noob ces hauv 48 tus neeg mob uas kuaj mob ADTKD. Cov neeg koom nrog lub raum tsis ua haujlwm los ntawm qhov tsis paub ua rau, nrog rau cov zis me me mus rau qhov tsis zoo. Cov neeg mob tau kuaj pom tias muaj ADTKD yog tias lawv muaj keeb kwm zoo hauv tsev neeg ntawm cov kab mob hauv lub raum ntev thiab / lossis hyperuricemia lossis yog tias cov kev tshawb pom ntawm lub raum biopsy tau qhia txog medullary cystic raum kab mob (lub sijhawm dhau los rau ADTKD-MUC1 thiab ADTKD-UMOD), tubulointerstitial nephritis, los yog mob interstitial puas. Peb tsis suav cov neeg mob uas muaj lub raum tsis zoo thiab/lossis cov tsos mob ntxiv. Cov qauv DNA tau kuaj thawj zaug siv SRS. Kev tshawb nrhiav ntxiv tau ua rau cov neeg mob uas tsis zoo SRS cov txiaj ntsig (n ¼ 39) siv LRS. Tag nrho cov qauv thiab cov ntaub ntawv kho mob tau txais los ntawm cov tsev kho mob xa mus rau Nyiv. Kev tshuaj ntsuam caj ces tau ua tom qab tau txais kev pom zoo los ntawm cov neeg mob thiab / lossis lawv cov neeg saib xyuas yog tias tus neeg mob yog menyuam yaus. Txoj kev tshawb no tau pom zoo los ntawm Lub Tsev Haujlwm Saib Xyuas Kev Tshawb Fawb ntawm Kobe University Tsev Kawm Ntawv Tshuaj Kho Mob (pom zoo 301).

2. Short-Read Sequencing

Genomic DNA was extracted from peripheral blood leukocytes using the QuickGene Mini 80 system (Wako Pure Chemical Industries, Ltd., Tokyo, Japan). For targeted sequencing using an SRS, samples were prepared using HaloPlex (Agilent Technologies, Santa Clara, CA) according to the manufacturer's instructions. Paired-end sequencing was performed on the MiSeq platform (Illumina, San Diego, CA). HaloPlex was used for targeted sequencing of 128 genes (version 2, Supplementary Table S1), 172 genes (version 4, Supplementary Table S2), 159 genes (version 5, Supplementary Table S3), 164 genes (version 6, Supplementary Table S4), 181 genes (version 7, Supplementary Table S5), and 183 genes (version 8, Supplementary Table S6) associated with congenital anomalies of the kidney and urinary tract, cystic kidneys, ADTKD, and nephronophthisis. The HaloPlex version used for each patient is found in Supplementary Table S7. Reads were aligned to the reference human genome (GRCh37/Hg19) using SureCall 4.0, which is a desktop application combining algorithms for end-to-end next-generation sequencing data analysis from alignment to categorization of mutations (Agilent Technologies). We excluded called variants with minor allele frequencies >1 feem pua ​​​​hauv cov ntaub ntawv sib txawv ntawm tib neeg, xws li cov hauv qab no: Human Genetic Variation Database (http://www.genome.med.kyoto-u.ac.jp/SnpDB/), gnomAD (https://gnomad.broadinstitute .org/), thiab 1000 Genomes Project (1000G) databases (https://www. internationalgenome.org/data). Cov neeg sib tw sib txawv tau raug xaiv los ntawm kev siv kev kwv yees software, SIFT (https://sift.bii.astar.edu.sg/), PolyPhen-2 (http://genetics.bwh.harvard.edu/pph2/) , Mutation Taster (http://www.mutationtaster.org/), thiab CADD (https://cadd.gs.washington.edu/snv), thiab tom qab ntawd muab faib ua cov kab mob, yuav muaj kab mob, lossis tsis paub tseeb, raws li cov Cov txheej txheem ntawm American College of Medical Genetics thiab Genomics.9

3. Kev Npaj Tsev Qiv Ntawv thiab Kev Nyeem Ntev Ntev (SMRT Sequence)

Cov amplicons tau sib sau ua ke hauv qhov sib npaug sib npaug thiab npaj rau SMRTbell cov tsev qiv ntawv sib txuas siv SMRTbell Express Template Prep Kit 2.0 (Pacific Biosciences, Menlo Park, CA) raws li cov chaw tsim khoom cov lus qhia. Tom qab ntawd, tus qauv ua ntu zus tau tsim los ntawm kev txuas cov txheej txheem primer version 2 thiab DNA polymerase rau cov adapters ntawm ob qhov kawg ntawm lub tsev qiv ntawv sib lawv liag, thiab cov qauv tau muab tso rau hauv SMRT cell. SMRT sequencing tau ua tiav siv PacBio Sequel II System nrog Sequel II Sequencing Kit 2.0. Cov ntaub ntawv ua ntu zus tau txheeb xyuas nrog SMRT Link version 9.0.0, thiab tau txais qhov kev pom zoo ua ntu zus. Tom qab ntawd, kev sib txuas ua ke tau ua los ntawm kev siv cov cuab yeej tshuaj ntsuam "pbaa," uas tsim nyog rau kev txheeb xyuas cov lus rov ua dua. Kev npaj tsev qiv ntawv, sib sau ua ke, thiab kev txheeb xyuas bioinformatics ntawm SMRT sequencing tau ua los ntawm Takara Bio (Kusatsu, Shiga, Nyiv). Cov nucleotide ib ntus, tus naj npawb ntawm kev nyeem, thiab allele zaus tau muab tso ua ke rau hauv Excel cov ntaub ntawv rau txhua tus neeg mob (cov ntaub ntawv tsis qhia). Rau txhua qhov kev txheeb xyuas, peb manually reconstructed lub sequence txhua txhua 60 lub hauv paus ib kab, raws li tau piav los ntawm Kirby li al.5 (Daim duab 1b). Vim tias muaj peev xwm PCR yuam kev hauv cov txheej txheem VNTR nyuaj, ntau qhov sib lawv liag tau kuaj pom hauv txhua tus neeg mob. Cov kab ke raug raug txheeb xyuas los ntawm kev sib piv cov txiaj ntsig LRS nrog cov electropherograms ntawm cov amplicons tau siv bioanalyzer (Table Ntxiv S8). Feem ntau, cov kab lus saum toj kawg nkaus 2 nrog cov neeg nyeem ntau tshaj plaws tau txiav txim siab kom raug. Qee qhov xwm txheej, txawm li cas los xij, cov kab ke nrog qhov thib peb lossis qis tshaj plaws ntawm cov ntawv nyeem tau txiav txim siab yog tias lawv qhov ntau thiab tsawg tau sib haum rau cov electrophoresis ncov pom los ntawm bioanalyzer (SC449, SC511, SC534, SC560, SC566, SC593, SC616, thiab SC639) vim qhov tsis muaj peev xwm ntawm PCR amplification ntawm ntev alleles thaum qhov loj ntawm VNTR cheeb tsam ntawm 2 alleles txawv heev.

Cistanche hmoov thiab Cistanche extract

Kev sib tham

Txhawm rau kom zoo tshaj plaws ntawm peb txoj kev paub, qhov no yog thawj daim ntawv tshaj tawm ntawm kev tshuaj ntsuam genetic tsom xam ntawm MUC1 VNTR variants kuaj pom siv kev sib xyaw ntawm SRS thiab LRS hauv ib pawg neeg Nyij Pooj. Peb tau txheeb xyuas MUC1 VNTR kev hloov pauv hauv 9 tus neeg mob los ntawm cov tsev neeg tsis sib xws. Tsis tas li ntawd, qhov kev kuaj pom tus nqi ntawm MUC1 qhov sib txawv ntawm cov neeg mob uas tau kuaj pom ADTKD yog 18.8 feem pua.

Raws li qhov nyuaj ntawm qhov sib txawv ntawm qhov hotspot, kuaj MUC1 VNTR qhov sib txawv los ntawm kev txheeb xyuas caj ces niaj hnub siv SRS yog qhov nyuaj, thiab lwm txoj hauv kev, xws li txoj kev snapshot lossis loj spectrometry, feem ntau yuav tsum tau. Hmoov tsis zoo, cov txheej txheem no yog qhov xav tau thiab tsuas yog ua tiav ntawm ob peb lub chaw soj nstuam thoob ntiaj teb.5,6 Wenzel thiab al.7 tau ua SMRT sib luag thiab lees paub txhua qhov kev kuaj mob hauv European cov neeg koom los ntawm 9 tsev neeg uas tau kuaj pom yav dhau los MUC1 VNTR sib txawv siv txoj kev snapshot. Wang et al.8 kuj tau qhia txog qhov muaj txiaj ntsig ntawm SMRT sequencing rau kev kuaj MUC1 VNTR kev hloov pauv hauv tsev neeg Suav loj. Txawm li cas los xij, vim lawv txoj kev tshawb fawb tsim, 2 qhov kev tshawb fawb no tsis tau txiav txim siab qhov kev kuaj pom ntawm MUC1 VNTR sib txawv hauv cov neeg mob uas muaj kev kuaj mob ADTKD. Hauv qhov kev tshawb fawb no, peb tau qhia meej tias qhov kev kuaj mob ntawm MUC1 qhov sib txawv hauv cov neeg mob uas kuaj pom tias ADTKD yog 18.8 feem pua, uas yuav pab tau kom muaj qhov tseeb ntau dua, tshwj xeeb tshaj yog thaum muaj kev sib tham txog caj ces.

Cov ntaub ntawv hais txog qhov muaj feem ntau ntawm ADTKD-MUC1 yog txwv, txawm hais tias cov kev tshawb fawb yav dhau los tau qhia txog qhov muaj ntau ntawm 0.7 mus rau 4 ib lab hauv Tebchaws Meskas thiab Ireland.3,13 Ib txoj kev tshawb fawb hauv tebchaws Askiv tau qhia txog kev kwv yees ntawm ADTKD. -UMOD ntawm 9 ib lab, tawm tswv yim ADTKD-UMOD yog cov kab mob hauv lub raum uas tsis muaj ntau tshaj plaws.14 Hauv peb txoj kev tshawb fawb, 9 cov neeg mob muaj MUC1 txawv, qhov 6 muaj UMOD variants, qhia ADTKD-MUC1 tuaj yeem yog 1 ntawm cov kab mob ntau tshaj plaws ntawm cov kab mob. ADTKD.

Txawm hais tias cov txheej txheem nyuaj ntawm MUC1 VNTR cheeb tsam, SRS tuaj yeem kuaj pom qhov txawv ntawm 3 tus neeg mob, txhua tus ntawm lawv muaj kev hloov pauv zoo ib yam: ib qho cytosine ntxig rau hauv 7 cytosine sib law liag ntawm qhov kawg ntawm 60-nucleotide chav nyob hauv qhov thib ob rov ua dua. ua VNTR. Nws tau xav tias qhov sib txawv hauv qhov ntiav ntawm MUC1 VNTR (kom txog rau thaum qhov thib peb rov ua dua ntawm VNTR) yuav tuaj yeem kuaj tau txawm tias DNA tau tawg (100 - 200 lub hauv paus ib khub nyeem) ua ib feem ntawm cov qauv kev npaj rau SRS tsom xam. Yamamoto et al.15 tau txheeb xyuas lwm qhov kev hloov pauv hloov pauv nyob ua ntej thaj tsam MUC1 VNTR siv SRS (tag nrho exome sequence). Qhov kev hloov pauv no tseem ua rau muaj qhov txawv txav ntawm cov protein nyob hauv tib yam li qhov kev hloov pauv nyob rau hauv VNTR.15 Cov txiaj ntsig no qhia tias SRS kev tshuaj ntsuam yuav muaj txiaj ntsig zoo rau kev txheeb xyuas qhov txawv ntawm qhov chaw ua ntej lossis hauv cheeb tsam ntiav ntawm VNTR.

Hauv peb txoj kev tshawb fawb, 7 ntawm 9 tus neeg mob nrog MUC1 VNTR variants muaj tib yam 60-nucleotide chav tsev ib ntus harboring cov cytosine ntxig (Table 2). Nyob rau hauv cov kev tshawb fawb yav dhau los uas tau nyeem ntev-nrhiav sequencing genetic tsom xam rau MUC1 VNTR tau ua, tib yam txawv tau kuaj pom nyob rau hauv 9 tsev neeg nyob rau hauv ib tug European cohort7 thiab ib tug loj Suav tsev neeg.8 Tsis tas li ntawd, qhov kev hloov no tau kuaj pom siv txoj kev snapshot thiab yog txhais raws li "27dupC." 12 Olinger li al.16 tau tshaj tawm tias qhov muaj feem ntau ntawm 27dupC variant yog 93.5 feem pua ​​​​hauv 2 qhov kev sau npe ntawm ADTKD hauv Tebchaws Europe thiab Tebchaws Meskas. Los ntawm cov kev tshawb pom no, peb cov ntaub ntawv tau qhia tias 27dupC variant yog qhov feem ntau MUC1 kev hloov pauv hauv ib pawg neeg Nyij Pooj.

Cistanche ntxiv

Tus naj npawb ntawm VNTR rov hais dua tau tshaj tawm tias yog 20 txog 253 ; Txawm li cas los xij, hauv peb txoj kev tshawb fawb, VNTR tus lej rov ua dua li ntawm 30 txog 82, nrog 80 feem pua ​​​​ntawm cov alleles muaj 30 txog 36 rov ua dua. Ib txoj kev tshawb fawb yav dhau los hauv Tebchaws Europe tau tshaj tawm tias feem pua ​​​​ntawm cov alleles nrog 30 txog 36 rov ua dua yog 39 feem pua ​​​​, uas tej zaum yuav raug txhawb los ntawm haiv neeg homogeneity ntawm cov neeg Nyij Pooj. Txoj kev tshawb fawb tam sim no tau qhia tias lub hnub nyoog nruab nrab ntawm kev tsim ESKD yog 45 xyoo (95 feem pua ​​​​CI: 40-50 xyoo) hauv cov neeg mob ADTKD-MUC1, suav nrog cov neeg mob hauv tsev neeg hauv pawg neeg Nyij Pooj (9 tsev neeg, n ¼ 31). Ib txoj kev tshawb fawb yav dhau los tau soj ntsuam 147 tus neeg, suav nrog cov neeg mob MUC1 kev hloov pauv thiab lawv cov neeg hauv tsev neeg cuam tshuam, thiab qhia tias lub hnub nyoog nruab nrab ntawm kev tsim ESKD yog 44.9 15.4 xyoo.17 Qhov sib txawv, kev tshawb fawb tsis ntev los no siv cov pab pawg sib txawv qhia tias lub sijhawm nruab nrab ntawm lub raum ciaj sia nyob hauv 104 cov neeg mob nrog ADTKD-MUC1 yog 36 xyoo (interquartile ntau yam: 30–46 xyoo), uas tej zaum yuav piav qhia los ntawm kev cais tawm ntawm cov neeg hauv tsev neeg uas tsis tau txais kev tshuaj ntsuam genetic tsom xam.16 Tsis tas li ntawd, cov kev hloov pauv ntawm interfamilial thiab intrafamilial tau raug tshaj tawm hauv cov neeg mob. nrog MUC1 variants, 8,17,18 txawm hais tias lub neej ntawm cov noob hloov pauv lossis lwm yam tseem tsis tau paub. Nyob rau tib lub sijhawm kawm, peb kuj tau kuaj pom kev hloov pauv hauv HNF1B, cov noob caj noob ces rau ADTKD, hauv 19 tus neeg mob. Txawm li cas los xij, cov neeg mob uas xav tias HNF1B kev hloov pauv tau raug tshem tawm los ntawm peb txoj kev tshawb fawb ntawm kev sau npe, vim tias feem ntau ntawm cov neeg mob no tau kuaj pom qhov txawv ntawm cov neeg muaj ADTKD-MUC1 vim muaj qhov txawv txav ntawm lub raum urinary system thiab / lossis autosomal-dominant tsev neeg keeb kwm ntawm. Thaum ntxov mob ntshav qab zib mellitus thiab / lossis hypomagnesemia, thiab peb tau tshaj tawm yav dhau los rau cov neeg mob no.19 Tseeb tiag, kab mob raum: Txhim kho cov txiaj ntsig thoob ntiaj teb kev pom zoo thiab kev tshuaj xyuas tsis ntev los no ntawm ADTKD tau tawm tswv yim tias lo lus ADTKD-HNF1B yuav tsum tsuas yog tshwj tseg rau cov xwm txheej hauv uas lub raum tubulointerstitial fibrosis yog qhov ua tshwm sim vim tsuas yog ob peb kis uas muaj tus kab mob tubulointerstitial nkaus xwb.1,20 Me nyuam yaus anemia, me hypotension, thiab me me hyperkalemia yog xav tias yog cov yam ntxwv ntawm ADTKD-REN. Txawm li cas los xij, kev tshawb fawb thoob ntiaj teb tsis ntev los no ntawm kev soj ntsuam cov yam ntxwv ntawm 111 ADTKD-REN cov neeg mob tau tshaj tawm tias kwv yees li 70 feem pua ​​​​ntawm cov neeg mob tau nthuav tawm rau cov tsev kho mob rau cov kab mob raum ntev lossis gout.21 Raws li tsab ntawv ceeb toom no, thaum yau anemia muaj nyob rau hauv 75.8 feem pua. cov neeg mob tab sis nws qhov mob hnyav heev; Qhov nruab nrab hemoglobin yog 9.6, 10.1, thiab 10.5 g / dl rau hnub nyoog<10 years, 10 to <15 years, and 15 to <20 years, respectively.21 Although hypotension was not reported in the article, the severity of hyperkalemia was mild and the mean serum potassium level in patients who were not taking fludrocortisone was 4.8 mEq/l.21 Childhood anemia, hypotension, or hyperkalemia may often be overlooked, and they are not always specific to ADRKD-REN; thus, we did not exclude participants with these findings. ADTKD-SEC61A1 is much rare than other ADTKD subtypes, with 6 families reported as of date.22 Patients harboring SEC61A1 mutation present with specific features, such as intrauterine growth retardation, cleft palate, congenital anemia, neutropenia, and immunodeficiency.3,22 Although the clinical manifestations of ADTKD-REN have not been completely clarified owing to the small number of reported cases, patients presenting with these specific features can be clearly distinguished from those with ADTKD-MUC1.

Cistanche tubulosa

Cov kev tshawb fawb tsis ntev los no ntawm cov txheej txheem molecular thiab cellular ntawm ADTKD-MUC1 tau qhia tias tus mob no yog ib qho tshuaj lom neeg muaj protein ntau tshwm sim los ntawm intracellular tsub zuj zuj ntawm misfolded MUC1 protein.11,23 Dvela-Levitt et al.24 qhia txog kev siv cov molecule me me, BRD4780. , txhawm rau hloov txoj hauv kev zais cia rau lysosomes, nthuav tawm MUC1fs raug tshem tawm hauv cov nas thiab cov neeg mob organoids thiab qhia txog cov peev xwm kho mob ntawm BRD4780. Yog li, kev kuaj pom tseeb ntawm MUC1 qhov sib txawv yuav yog qhov tseem ceeb rau kev txhim kho cov tshuaj kho tau zoo. Peb txoj kev tshawb fawb muaj ntau yam kev txwv. Ua ntej, tag nrho peb cov neeg koom nrog kev kawm yog neeg Nyij Pooj, thiab peb cov qauv loj me me vim lub sijhawm kawm luv. Muab qhov xwm txheej rov qab los ntawm txoj kev tshawb no, peb kuj tsis tuaj yeem tau txais cov ntaub ntawv kho mob ntev ntev, xws li mob raum. Qhov thib ob, ib leeg-base insertions tau pom txawm nyob rau hauv alleles tsis muaj kev hloov. Raws li qhov ua yuam kev siab, nws nyuaj kom ua tiav kev sib dhos zoo kom pom ib leeg-nucleotide variants lossis indels siv LRS. Nws yog qhov tsim nyog los sib xyaw ua ntu zus los ntes txhua hom kev sib txawv ntawm caj ces, uas nce tus nqi thiab kev nyuaj siab ntawm cov phiaj xwm. Yog li, PacBio tau tsim ib qho kev sib txuas tshiab, lub npe hu ua voj voog kev pom zoo ua ke. Lub voj voog kev pom zoo sib sau ua ke ua rau muaj kev ntseeg siab nyeem nrog 99.8 feem pua ​​​​qhov raug thiab qhov nruab nrab ntev ntawm 13.5 kilobase, uas ua rau txo qis qhov kev ua yuam kev ua ntu zus.25 Txoj kev sib txuas no kuj tau siv hauv peb txoj kev tshawb fawb. Yog li ntawd, txiav txim siab txog kev txhim kho hauv qhov kev ua tiav qhov tseeb, ib qho kev ntxig ib leeg tuaj yeem tshwm sim los ntawm PCR yuam kev. Txawm hais tias KOD Kub Pib DNA polymerase tau ua kom zoo rau kev nthuav dav ntawm cov hom phiaj nyuaj tshaj plaws, 26 lwm cov DNA polymerases, qhov kev ncaj ncees uas siab dua li ntawm KOD (xws li Phusion), tuaj yeem pab txo qis PCR qhov yuam kev.27 Tsis ntev los no cov kev tshawb fawb tau pom ib qho amplification-free raws tu qauv rau lub hom phiaj enrichment siv cov clustered tsis tu ncua interspaced luv palindromic repeats/Cas9 system.28,29 Txoj kev tshiab no yuav muab tau zoo dua. Thib peb, peb tsis tau lees paub lwm txoj hauv kev uas tuaj yeem pab kuaj mob, xws li kev tiv thaiv kab mob raum lossis cov qe ntshav los ntawm cov zis. Kirby et al., 5 uas thawj zaug tshaj tawm qhov kev hloov pauv rau ADTKD-MUC1, tau ua qhov kev tshuaj ntsuam xyuas immunohistochemical ntawm lub raum kuaj los ntawm cov neeg mob MUC1 kev hloov pauv. Lawv tau tshaj tawm tias, txawm hais tias kev tiv thaiv kab mob hauv ib txwm muaj ua rau tsis muaj qhov tshwj xeeb, cov neeg mob MUC1 kev hloov pauv tau nthuav tawm cov qauv tshwj xeeb hauv cov kab mob hauv lub voj voog ntawm Henle, cov tubules distal, thiab sau cov ducts.5 Cov kev soj ntsuam zoo sib xws ntawm cov mutant MUC1 protein tau raug tshaj tawm. 11,13,18,30 Yamamoto thiab al.15 kuaj pom mutant MUC1 protein nyob rau hauv cov zis exosomes. Txawm hais tias qhov kev kho mob tshwm sim ntawm ADTKD-MUC1 tsis yog qhov tshwj xeeb, cov kev tshuaj ntsuam tsis muaj noob caj noob ces tuaj yeem pab txhawb kev kuaj mob. Yog li, nws yog ib qho tsim nyog los txheeb xyuas qhov muaj txiaj ntsig ntawm cov kev tshuaj ntsuam tsis muaj noob caj noob ces ua ke nrog cov txiaj ntsig ntawm peb txoj kev tshawb fawb. Hauv kev xaus, peb txoj kev tshawb fawb qhia tau hais tias kev sib txuas ntawm caj ces nrog SRS thiab LRS yog qhov muaj txiaj ntsig zoo rau kev tshawb nrhiav MUC1 VNTR sib txawv hauv ib pawg neeg Nyij Pooj thiab tias qhov muaj feem ntau ntawm ADTKD-MUC1 yuav muaj ntau hauv cov pejxeem. Kev sib xyaw ntawm kev tshuaj ntsuam caj ces siv SRS thiab LRS zoo li yuav txhim kho qhov kev kuaj mob ntawm ADTKD-MUC1 thiab tuaj yeem pab txhawb kev txhim kho kev kho kom zoo.

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Eri Okada1,2 , Naoya Morisada1,3 , Tomoko Horinouchi1 , Hideki Fujii4 , Takayuki Tsuji5 , Masayoshi Miura6 , Hideyuki Katori7 , Masashi Kitagawa8 , Kunio Morozumi9 , Takanobu Toriyama10 , Yuki11Komi 2 Nakamura , sushi Kondo1, Yuya Aoto1, Shinya Ishiko1, Rini Rossanti1, Nana Sakakibara1, Tuam Tshoj Nagano1, Tomohiko Yamamura1, Shingo Ishimori1, Joichi Usui2, Kunihiro Yamagata2, Kazumoto Iijima13,14, Toshiyuki Imasawa15 thiab Kandai Nozu1