MicroRNAs nyob rau hauv lub raum Kev Loj Hlob thiab Kab Mob Ⅲ

MiRNAs hauv Wilms qog

Wilms qog,los yogmob nephroblastoma, yog cov menyuam yaus feem ntaumob qog noj ntshav, nrog qhov tshwm sim ntawm 1 ntawm 10, 000 cov menyuam yaus hauv North America (111). Nws feem ntau yog kab mob sib kis, txawm hais tias tsev neeg cov ntaub ntawv tshwm sim hauv kwv yees li 1% -2% ntawm cov neeg mob (112, 113). Wilms hlav tshwm sim los ntawm aberrant nephrogenesis, qhov twg pluripotent embryonic lub raum precursors tsis sib txawv thiab pheej.txawv txav mus rau hauv lub neej postnatal(111, 114, 115). Cov qog no histological zoo litsim lub raumnrog acuam ​​tshuam morphology(116), thiab kev hloov pauv hauv cov noob koom nrog hauv fetal nephrogenesis, suav nrog WT1 (117–119), CTNNB1 (120), SIX1/2 (121, 122), thiab TP53 (123, 124), muaj feem xyuam nrog kwv yees li 40% ntawm Wilms qog (125). Cov kev tshawb fawb tsis ntev los no siv tag nrho-genome thiab tag nrho-exome sequencing ntawm Wilms cov qog pom tau tias muaj kev hloov pauv tshiab hauv cov noob qog noj ntshav (REST, CHEK2, PALB2) (126, 127), cov noob encoding proteins uas kho cov kev hloov kho histone thaum lub sij hawm nephrogenesis (BCOR, MAP3K4) (126. ), thiab miRNA-processing genes (121, 122, 128, 129).

YUAV UA LI CAS RAU CISTANCHE ua haujlwm rau cov neeg mob rau lub raum?

Ntawm cov miRNA-processing genes, kev hloov pauv hauv DROSHA, DGCR8, DICER1, TARBP2, thiab XPO5 (encodes exportin 5) tau tshaj tawm hauv kev kho mob-naive thiab neoadjuvant chemotherapy-kho Wilms qog (Daim duab 3) (121, 128, 121, 121, 12, 12, 121, 121, 121, 121, 121, 121, ). Kwv yees li 33% ntawm Wilms qog kuaj pom muaj kev hloov pauv tsis zoo hauv cov noob ntawm miRNA-txhim kho txoj hauv kev (128). Ib qho kev hloov pauv hotspot rov tshwm sim (E1147K) hauv kev sib ntsib al-binding (Mg2+) residue ntawm RNase IIIb domain ntawm DROSHA, uas zoo nkaus li tshwj xeeb rau Wilms qog, tshem tawm cov haujlwm catalytic ntawm lub npe no, ua rau tsis tiav. cleavage ntawm pri-miRNAs thiab txo miRNA maturation (128). Somatic hotspot kev hloov pauv cuam tshuam rau RNase IIIb ntawm DICER1 cuam tshuam kev ua haujlwm ntawm 5p miRNAs (cov uas tau los ntawm 5'-arm ntawm pre-miRNA hairpin) (129) thiab feem ntau pom tias yog "thib ob" kev hloov pauv uas ua nyob rau hauv tandem nrog DICER1 germline hloov mus rau induce Wilms tumorigenesis nyob rau hauv DICER1 syndrome (ib qho teeb meem uas ua rau kom muaj ntau yam qog nqaij hlav) (130-132). Nws tseem tsis tau paub meej tias vim li cas DICER1 kev ua haujlwm tsis zoo hauv Wilms cov qog ua rau muaj kev pheej hmoo thiab tsis meej pem nephrogenesis, tsis zoo li qhov poob ntawm Dicer1 hauv nas nephron progenitors, uas ua rau muaj kev nce apoptosis thiab ua rau ntxov ntxov ntawm nephrogenesis (72-75). Qee qhov muaj peev xwm muaj peev xwm suav nrog qhov kev siv tshuaj ntau npaum li cas yuav yog qhov tseem ceeb hauv kev txiav txim siab ntawm cell ciaj sia lossis qhov hloov pauv hauv RNase IIIb tus thawj coj yuav cuam tshuam rau qhov tshwj xeeb ntawm miRNA khi los ntawm DICER1.

Kev hloov pauv hauv miRNA-processing genes cuam tshuam nrog kev txo qis ntawm cov miRNA tseem ceeb, suav nrog cov tswv cuab ntawm miR-200 (121) thiab let-7 tsev neeg (Daim duab 3) (121, 129). Cia-7 miRNAs thiab RNA-binding protein Lin28 muaj nuj nqi hauv kev hais kwv txhiaj los tswj lub sijhawm ntawm kev tso tseg ntawm murine nephrogenesis, tejzaum nws yog los ntawm kev tswj hwm ntawm kev loj hlob ntawm cov noob Igf2 (133). Overexpression ntawm Lin28 thaum lub raum kev loj hlob ua rau expansion ntawm nephrogenic progenitors, los ntawm inhibiting lawv kawg nthwv dej ntawm kev sib txawv, uas culminates nyob rau hauv neoplastic transformation uas yog heev reminiscent ntawm tib neeg Wilms qog (134). Ntxiv DNA luam tus naj npawb ntawm LIN28B thiab DNA daim ntawv poob ntawm let{16}}a yog pom nyob rau hauv 25% thiab 46% ntawm tib neeg Wilms qog qog, raws li (126). Nyob rau hauv txoj kab nrog cov kev soj ntsuam no, germline hloov pauv hauv tib neeg DISL3L2 noob, uas encodes ib qho exoribonuclease lub luag hauj lwm rau degrading preprocessed daim ntawv ntawm let-7, ua rau Perlman syndrome thiab predisposition rau Wilms qog (135, 136). Perlman Syndrome yog ib tug congenital overgrowth syndrome tshwm sim los ntawm macrosomia, polyhydramnios, lub ntsej muag dysmorphology, lub raum dysplasia, thiab nephroblastomatosis (ib qho mob ua ntej rau Wilms qog) (137). Ntawm cov menyuam mos uas muaj Perlman syndrome uas muaj sia nyob dhau lub sijhawm yug menyuam, 64% tsim Wilms qog (138). Interestingly, ua kom tiav los yog ib nrab DISL3L2 tshem tawm tau pom nyob rau hauv kwv yees li 30% ntawm cov qog nqaij hlav Wilms kuaj (136).

Ib txoj kev tshawb fawb tsis ntev los no tau ntxiv dag zog rau qhov tseem ceeb ntawm miRNA kev tswj hwm network hauv etiology ntawm Wilms qog. Cov kws sau ntawv tau pom tias pleiomorphic adenoma gene 1 (PLAG1) yog ib qho ntawm cov noob caj noob ces nyob rau hauv Wilms qog nrog kev hloov pauv hauv miRNA-processing genes (125). Ectopic qhia ntawm PLAG1 nyob rau hauv lub tsim nas lub raum ua rau neoplasia, uas yog nrog los ntawm transactivation ntawm nws lub hom phiaj noob, lub Wilms qog oncogene IGF2. miR-16 thiab miR-34, uas yog downregulated nyob rau hauv Wilms qog, raug txheeb xyuas tias muaj peev xwm regulators ntawm PLAG1 qhia (125). Table 2 piav qhia txog lwm cov kev tshawb fawb uas tau tshaj tawm kev qhia tsis meej ntawm cov miRNAs tshwj xeeb cuam tshuam nrog etiology ntawm Wilms qog. Qhov zoo siab, cov miRNAs tuaj yeem ua haujlwm raws li oncogenes (hu ua oncomiRs) lossis cov qog nqaij hlav hauv qhov chaw ntawm Wilms qog kev loj hlob, nyob ntawm seb lawv lub hom phiaj.

MiRNAs raws li biomarkers muaj peev xwm thiab cov kws kho mob

Sib nrug los ntawm lawv qhov chaw nyob hauv lub cev, miRNAs kuj tseem muaj nyob hauv cov kua roj lom, suav nrog ntshav, ntshav, zis, mis, thiab qaub ncaug (139). Cov miRNA circulating no pom tau ntim cov hnub nyoog hauv microparticles (exosomes, microvesicles, thiab apoptotic cev) (140, 141), conjugated nrog AGO (142) los yog nucleophosmin 1 proteins (143), los yog loaded rau hauv HDL (144), uas ua rau lawv zoo kawg li. ruaj khov txawm tias nyob rau hauv cov xwm txheej tsis zoo, xws li boiling, huab variations nyob rau hauv pH, ncua cia, thiab ntau yam khov-thaw mus (145, 146). Yog li, miRNA kos npe rau hauv cov kua roj ntsha tuaj yeem cuam tshuam txog kev koom tes nrog lub cev lossis kab mob (147). Ua ke, cov yam ntxwv no ua rau kev nthuav dav miRNAs txaus nyiam siv los ua cov cim biomarkers tsis muaj kab mob rau kev kuaj mob thiab kev kuaj mob.

Circulating miRNAs tuaj yeem muab rho tawm ncaj qha los ntawm cov kua roj ntsha uas tsis muaj kev cuam tshuam los yog cov khoom siv ntxiv ntawm lar vesicle uas siv cov khoom siv rho tawm (148) lossis TRIzol (149). Thaum kev sib cais, miRNAs tuaj yeem khaws cia ntawm -70 degree thiab nyob ruaj khov mus txog 1 xyoo (148). Muaj ntau lub platforms muaj rau miRNA profileing, suav nrog microarray hybridization, qPCR, thiab lwm tiam kev ua sequencing (150). Microarray thiab qPCR yog cov txheej txheem uas nquag siv los tshawb xyuas qhov qhia ntawm miRNAs (151). Ob txoj hau kev muaj qhov zoo ntawm kev siv yooj yim, kuj ceev los ntawm RNA sau npe rau cov ntaub ntawv tsim, thiab tus nqi zoo (152). Txawm li cas los xij, lawv cia siab rau qhov muaj thiab cov lus qhia tseeb ntawm miRNA ib ntus hauv cov ntaub ntawv rau kev sojntsuam thiab primer tsim (150). Txawm hais tias kim dua, tiam tom ntej txuas ntxiv tso cai rau kev tshawb nrhiav ib txhij ntawm ob qho tib si paub thiab tshiab miRNA hom thiab muaj kev nkag siab zoo (153). Tsis tas li ntawd, qhov kev daws teeb meem ib leeg-nucleotide ntawm lwm tiam tom ntej ua rau muaj kev txheeb xyuas ntawm isomiRs, uas yog mRNA isoforms uas paub tab uas txawv ntawm cov canonical ntev, sib lawv liag, lossis ob qho tib si (154, 155), uas hloov lub hom phiaj ntawm miRNA ( 156).

Cov kev tshawb fawb sib txawv tau tshawb xyuas lub peev xwm ntawm kev nthuav tawm miRNAs ua biomarkers rau cov kab mob hauv lub raum (157-160). Piv txwv li, ib txoj kev tshawb fawb tau txheeb xyuas 14 miRNAs uas tau hloov kho hauv cov ntshav ntawm cov neeg mob Wilms qog. Qhov zoo siab, kos npe raws li miR-100-5p thiab miR-130-3p kev qhia tuaj yeem ua rau cov neeg mob sib txawv ntawm kev tswj hwm kev noj qab haus huv nrog qhov tseeb, rhiab heev, thiab tshwj xeeb (159). Txawm hais tias qhov kev tshawb pom los ntawm qhov kev tshawb fawb no thiab ntau lwm cov kev tshawb fawb tau muab kev txhawb zog los tshawb txog lub peev xwm ntawm kev nthuav dav miRNAs ua biomarkers, ntau qhov teeb meem hauv cheeb tsam yuav tsum tau kov yeej ua ntej kev siv tshuaj kho mob thoob plaws. Ua ntej, muaj cov txheeb ze tsis muaj kev pom zoo ntawm cov kev tshawb fawb yuav tshwm sim vim tsis muaj cov txheej txheem kev ua kom huv (161) thiab kev soj ntsuam ntawm cov qauv (xws li, qhov sib txawv hauv miRNA profileing platforms, refs. 162, 163; los yog qhov sib txawv hauv smRNA-Seq tsev qiv ntawv kev npaj, ref. 164). Qhov thib ob, yog qhov tsis muaj cov qauv loj-loj cov kev tshawb fawb thiab cov ncauj lus kom ntxaws txog kev tshawb fawb tshwj xeebkab mob.Lwm qhov tseem ceeb yog qhov cuam tshuam ntawm qhov tsis sib haum xeeb xws li hnub nyoog, poj niam txiv neej, thiab lwm yam (xws li, luam yeeb, cawv, thiab lwm yam) ntawm miRNA profiles tsis tau tshawb pom tag nrho (rau kev tshuaj xyuas qhov tob, thov xa mus rau ref. 165).

Ntawm cov kws kho mob, ntau cov tshuaj siv miRNA tam sim no nyob rau hauv kev sim tshuaj tab sis tseem tsis tau txais kev pom zoo los ntawm FDA (166, 167). Txoj hauv kev tseem ceeb rau kev kho miRNA suav nrog kev kho dua ntawm miRNA qib siv miRNA mimics, lossis inhibition ntawm miRNAs tshwj xeeb siv antagomiRs (168). Ib qho ntawm cov teeb meem cuam tshuam nrog kev txhim kho ntawm miRNA-raws li kev kho mob yog kev txheeb xyuas cov neeg sib tw miRNA rau txhua tus kab mob. Vim tias muaj ntau yam miRNAs tsis zoo nyob rau hauv txhua tus kab mob, ua tib zoo tshuaj xyuas ntawm tus neeg mob cov qauv ua ke nrog hauv vitro thiab hauv vivo kev soj ntsuam uas hais txog cov txheej txheem pathophysiological cuam tshuam los ntawm miRNAs hauv nqe lus nug yuav tsum tau ua kom txo qis tus neeg sib tw miRNAs rau kev kho mob (168, 169). Lwm qhov kev sib tw muaj xws li kev txhim kho cov tswv yim los txhim kho hauv vivo kev ruaj ntseg thiab qhov chaw tshwj xeeb cov tshuaj xa nrog tsawg kawg nkaus toxicity thiab off-target los. RNA molecules yog tshuaj tsis ruaj khov vim muaj 2'-hydroxyl pawg ntawm lub nplhaib pentose. Txhawm rau muab kev ruaj ntseg dua thiab kev tiv thaiv los ntawm nucleases tam sim no nyob rau hauv cov ntshav los yog cov endocytic compartment ntawm cov hlwb, biotech tuam txhab uas muag tau tsim RNA molecules nrog tshuaj hloov kho (2′-O-methyl pawg, phosphorothioate, los yog kaw nucleic acids) nyob rau hauv lawv cov qaum (166). Raws li rau hauv vivo kev xa khoom, thev naus laus zis suav nrog lipid-based (piv txwv li, lipid nanoparticle thiab nruab nrab liposome) thiab dendrimers conjugated rau lub hom phiaj moiety, ntawm ntau lwm cov tswv yim. Cov teeb meem loj cuam tshuam nrog miRNA cov tshuab xa khoom yog kev tiv thaiv kab mob thiab cov phiaj xwm tshwj xeeb rau cov kab mob (170). Kev xa tawm cov tswv yim los ntawm ntau txoj kev tswj hwm (intraperitoneal, intravenous, thiab subcutaneous txhaj) los yog los ntawm kev siv cov vectors uas muaj raum tshwj xeeb thiab inducible txhawb tau zoo siv rau kev xaiv lub raum tsom thiab kom tsis txhob muaj kev cuam tshuam rau lwm cov ntaub so ntswg thiab kab mob (171, 172. ).

Table 2. Cov npe ntawm miRNAs hloov pauv hauv Wilms qog

Cov npe ntawm miRNAs, cov qauv / cov qauv tshuaj ntsuam xyuas, qib miRNA tau pom nyob rau hauv cov qauv no / cov qauv piv nrog cov kev tswj kom tsim nyog, lub hom phiaj gene, qhov tshwm sim ntawm miRNA kev hloov pauv, thiab cov ntaub ntawv pov thawj. CAMKK2, calcium / calmodulin-dependent protein kinase kinase 2; CDC7, cell faib voj voog 7; CDH1, cadherin 1; CREB1, cAMP- teb cov ntsiab lus-binding protein 1; EYA1, qhov muag tsis pom homolog 1; E2F3, E2F transcription factor 3; FRS2, FGF receptor substrate 2; Glut1, glucose transporter 1; IGF1R, IGF1 receptor; Yog 1, jagged1; MET, kev hloov mesenchymal-epithelial; MKNK1, MAPK-interacting serine/threonine kinase 1; PTEN, phosphatase thiab tensin homolog; PUMA, p53-upregulated modulator ntawm apoptosis; p73, qog protein p73; SIX1, sineoculis homeobox homolog 1; TGFBR1, TGF- receptor 1; WT, Wilms qog.

Cov ntsiab lus

Muaj kev tawg ntawm cov ntaub ntawv hais txog miRNA biogenesis, kev cai ntawm miRNA qhia, thiab miRNA muaj nuj nqi txij li thaum pib nrhiav pom miRNAs hauv 1993 (6, 7). Qhov no tau nrog los ntawm kev nkag siab ntau ntxiv ntawm miRNAs ua haujlwm li cas ob qho tib si hauv physiology thiab hauv pathophysiology ntawm ntau yam kab mob. Nws tau dhau los ua qhov tseeb tias dysregulation ntawm miRNA qhia cuam tshuam rau kev loj hlob ntawm lub raum thaum ntxov thiab cuam tshuam rau hauv cov kab mob ntawm cov kab mob hauv lub raum, xws li CAKUT thiab Wilms qog. Nrog rau kev txhim kho tsis ntev los no hauv kev siv miRNAs ua biomarkers thiab ua cov hom phiaj tshuaj tshiab, kev nkag siab txog yuav ua li cas miRNAs tswj lub raum kev loj hlob thiab kab mob yog qhov tseem ceeb kom nkag siab txog yuav ua li cas lawv thiaj li yuav siv tau rau hauv kev kuaj mob tshiab thiab kho cov kab mob no. Txhawm rau kom paub txog cov kev siv zog no, cov kev tshawb fawb yav tom ntej txheeb xyuas qhov kev ua haujlwm ntawm miRNAs tshwj xeeb hauv kev loj hlob ntawm lub raum yog qhov tseem ceeb, ntxiv rau cov thev naus laus zis los txhim kho lub hom phiaj me me oligonucleotide kho rau lub raum.

Kev lees paub

JH lub chaw kuaj mob tau txais kev txhawb nqa los ntawm NIH National Institute of Diabetes thiab Digestive and Kidney Diseases (R01DK125015 thiab R01DK102843). DMC tau txais kev txhawb nqa los ntawm Nephrotic Syndrome Study Network Career Development Award thiab Children's Hospital of Pittsburgh Research Advisory Council Postdoctoral Fellowship. MT tau txais kev txhawb nqa los ntawm University of Pittsburgh Summer Research Internship Program (R25DK119180). Cov duab tau tsim nrog BioRender.com. Chaw nyob: Jacqueline Ho, Division of Nephrology, Department of Pediatrics, UPMC Children's Hospital of Pittsburgh, Rangos Research Center, 4401 Penn Ave., Pittsburgh, Pennsylvania 15224, USA. Xov tooj: 412.692.9440; Email: jacqueline.ho2@chp.edu.

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