DOCSLIB.ORG

NPHS2 Gene Mutation, Atopy, and Gender As Risk Factors for Steroid-Resistant Nephrotic Syndrome in Indonesians

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
NPHS2 Gene Mutation, Atopy, and Gender As Risk Factors for Steroid-Resistant Nephrotic Syndrome in Indonesians Paediatrica Indonesiana VOLUME 51 September NUMBER 5 Original Article NPHS2 gene mutation, atopy, and gender as risk factors for steroid-resistant nephrotic syndrome in Indonesians Dedi Rachmadi, Dany Hilmanto, Ponpon Idjradinata, Abdurahman Sukadi Abstract QUHFHQW\HDUVPROHFXODUJHQHWLFVWXGLHVKDYH Background 6WHURLGUHVLVWDQWQHSKURWLFV\QGURPH 6516 RIWHQ EHHQZHOOGHYHORSHGLQFOXGLQJVWXGLHVIRU GHYHORSVLQWRHQGVWDJHUHQDOGLVHDVH3UHYLRXVVWXGLHVKDYH JHQHVLQYROYHGLQWKHSDWKRJHQHVLVRIQHSKURWLF reported that NPHS2JHQHPXWDWLRQJHQGHUDQGDWRSLFKLVWRU\ V\QGURPH2ISDUWLFXODULQWHUHVWLVWKHJHQH DUHULVNIDFWRUVDVVRFLDWHGZLWK6516,QWHUHWKQLFVRFLRFXOWXUDO I DQGHQYLURQPHQWDOGLIIHUHQFHVKDYHDOVREHHQVXJJHVWHGWRDIIHFW encoding a protein that maintains the diaphragm slit WKHVHPXWDWLRQV RISRGRF\WHV*HQHPXWDWLRQDQDO\VLVLVLPSRUWDQW Objective7RDQDO\]HSRVVLEOHULVNIDFWRUVIRU6516LQFOXGLQJ with regards to phenotype and predicting the severity NPHS2JHQHPXWDWLRQV &o7DQGGHO* JHQGHUDQG RIFOLQLFDOPDQLIHVWDWLRQV The NPHS2 gene encodes DWRSLFKLVWRU\LQ,QGRQHVLDQVXEMHFWVZLWK6516 the podocin protein and is located on chromosome Methods$FDVHFRQWUROVWXG\ZLWKVXEMHFWVFRQVLVWLQJRI 6516SDWLHQWVDQGFRQWUROVXEMHFWVZDVXQGHUWDNHQLQ T0XWDWLRQRIWKLVJHQHKDVEHHQDVVRFLDWHG ,QGRQHVLDQWHDFKLQJFHQWUHKRVSLWDOVIURP6HSWHPEHUWR ZLWKSRRUVWHURLGUHVSRQVHLQWUHDWPHQWRIQHSKURWLF 'HFHPEHU$QDO\VLVRIWKHNPHS2JHQHPXWDWLRQLQ V\QGURPH 16 3UHYLRXVVWXGLHVKDYHVKRZQWKDW &o7ZDVSHUIRUPHGE\DPSOLILFDWLRQUHIUDFWRU\PXWDWLRQV\VWHP SRNS patients with an NPHS2JHQHPXWDWLRQKDG SRO\PHUDVHFKDLQUHDFWLRQ $5063&5 ZKLOHWKDWIRUWKH NPHS2JHQHPXWDWLRQLQGHO*ZDVSHUIRUPHGE\UHVWULFWLRQ PRUHVHYHUHFOLQLFDOPDQLIHVWDWLRQVFRPSDUHGWR IUDJPHQWOHQJWKSRO\PRUSKLVP 5)/3 'DWDZDVDQDO\]HGE\ 6516SDWLHQWVZLWKRXWWKHNPHS2JHQHPXWDWLRQ PXOWLSOHORJLVWLFUHJUHVVLRQ NPHS2JHQHPXWDWLRQVKDYHEHHQIRXQGLQ Results,QRXU,QGRQHVLDQVXEMHFWVWKHVLJQLILFDQWULVNIDFWRUV RI6516SDWLHQWVPRVWO\SUHVHQWLQWKHWKLUGH[RQ IRU6516ZHUHPDOHJHQGHU 25 &, 5 3 NPHS2&o7JHQHPXWDWLRQ 25 &, RIWKHJHQH 2WKHUVWXGLHVRI6516SDWLHQWVDOVR 3 DQGNPHS2GHO*JHQHPXWDWLRQ VKRZHGPXWDWLRQVORFDWHGRQWKHWKLUGH[RQQDPHO\ 25 &,3 +RZHYHUDWRSLF DQRQVHQVHPXWDWLRQDW&o7 5; and a KLVWRU\ZDVQRWDVLJQLILFDQWULVNIDFWRUIRU6516 25 GHOHWLRQDWGHO*6 &,3 Conclusion NPHS2 &o7DQGGHO*JHQHPXWDWLRQV DVZHOODVPDOHJHQGHUDUHULVNIDFWRUVIRU6516LQ,QGRQHVLDQ VXEMHFWV$WRSLFKLVWRU\ZDVQRWVLJQLILFDQWO\DVVRFLDWHGZLWK6516 LQRXUVXEMHFWV[Paediatr Indones. 2011;51:272-6]. )URP'HSDUWPHQWRI&KLOG+HDOWK)DFXOW\RI0HGLFLQH3DGMDGMDUDQ 8QLYHUVLW\+DVDQ6DGLNLQ+RVSLWDO%DQGXQJ,QGRQHVLD Keywords: Steroid-resistant nephrotic syndrome, risk factor, NPHS2 gene mutation Reprint requests to:'HGL5DFKPDGL0''HSDUWPHQWRI&KLOG+HDOWK )DFXOW\RI0HGLFLQH3DGMDGMDUDQ8QLYHUVLW\+DVDQ6DGLNLQ+RVSLWDO-O 3DVWHXU%DQGXQJ7HO(PDLOGHGLUDFKPDGL# yahoo.com 272Paediatr Indones, Vol. 51, No. 5, September 2011 'HGL5DFKPDGLHWDO13+6JHQHPXWDWLRQDWRS\DQGJHQGHUDVULVNIDFWRUVIRU6516 Diversity in the NPHS2JHQHPXWDWLRQSDWWHUQ RIWKH3DGMDGMDUDQ8QLYHUVLW\0HGLFDO6FKRRO+DVDQ LVWKRXJKWWREHGXHWRYDULDWLRQVLQHWKQLFLW\DQG 6DGLNLQ*HQHUDO+RVSLWDO%DQGXQJ environment.6LQFHWKHUHZDVQRVWXGLHVRQ NS was diagnosed according to the International NPHS2JHQHPXWDWLRQVLQWKHSHGLDWULF,QGRQHVLDQ 6WXG\RI.LGQH\'LVHDVHLQ&KLOGUHQ ,6.'& FULWHULD SRSXODWLRQZHIRFXVHGRQWKHWKLUGH[RQRINPHS2 LHHGHPDVHYHUHSURWHLQXULDK\SRDOEXPLQHPLD DVDEDVLVIRUVWXG\LQ,QGRQHVLDQFKLOGUHQZLWK6516 JG/ ZKLOH6516ZDVGHILQHGDV16SDWLHQWV 6LQFHPDQ\IDFWRUVPD\SOD\DUROHLQWKHRFFXUUHQFH QRWDFKLHYLQJUHPLVVLRQDIWHUVLQJOHGUXJSUHGQLVRQH RI6516ZHLQFOXGHGJHQGHUDQGKLVWRU\RIDWRS\DV WKHUDS\XVLQJWKHIXOOGRVHIRUWKHILUVWIRXUZHHNV SRVVLEOHULVNIDFWRUVRI6516LQWKLVVWXG\2XUDLP NS diagnoses were made by pediatric nephrologists or ZDVWRGHWHUPLQHLINPHS2JHQHPXWDWLRQVJHQGHU SHGLDWULFLDQVVXSHUYLVHGE\SHGLDWULFQHSKURORJLVWV DQGKLVWRU\RIDWRS\DUHULVNIDFWRUVIRU6516LQ 'DWDDQDO\VHVZHUHSHUIRUPHGXVLQJ6366 Indonesian children. YHUVLRQ0XOWLSOHORJLVWLFUHJUHVVLRQDQDO\VLV ZDVXVHGWRGHWHUPLQHSRVVLEOHULVNIDFWRUVIRU6516 LQFOXGLQJJHQHPXWDWLRQV NPHS2&oT and Methods GHO*JHQGHUDQGIDPLOLDODWRSLFKLVWRU\ To detect the NPHS2&o7PXWDWLRQLQ $FDVHFRQWUROVWXG\ZDVFRQGXFWHGIURP6HSWHPEHU H[RQZHXVHG$5063&53ULPHUGHVLJQLVVKRZQ WR'HFHPEHU:HLQFOXGHGFDVHVXEMHFWV in Figure 16XEMHFWV·'1$VSHFLPHQVZHUHSODFHG with primary SRNS aged between 1-14 years old. LQGXSOLFDWHWXEHVRQHWXEHFRQWDLQLQJWKHZLOGW\SH &RQWUROVXEMHFWVZHUHKHDOWK\FKLOGUHQZLWKQR DQGUHYHUVHSULPHUVDQGWKHRWKHUWXEHFRQWDLQLQJ NLGQH\GLVHDVHVRUFRQJHQLWDODEQRUPDOLWLHV&DVH WKHPXWDWHGDQGUHYHUVHSULPHUV6LQFHNPHS2 is an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··**77*7$&$$*$*7$$7**$$$*$*7$$77$7$77$& 0XWDWHG 7 SULPHU··**77*7$&$$*$*7$$7**$$$*$*7$$77$7$77$T 5HYHUVHSULPHU··7*$$*$$$77**&$$*7&$* Figure 1. Wild type sequence, mutated sequence and reverse primers for ARMS-PCR. T **77*7$&$$*$*7$$7**$$$*$*7$$77$7$77$&*$&7***$&$7&&*&77 &&7**$$*$*&&$$$**&&&7**7$$$$$$$&$&7&77777777&7$$$&$&&7& 7&7&&7*$&77*&&$$777&77&$ Figure 2. Predicted PCR product, 131 base pairs. Paediatr Indones, Vol. 51, No. 5, September 2011 273 'HGL5DFKPDGLHWDO13+6JHQHPXWDWLRQDWRS\DQGJHQGHUDVULVNIDFWRUVIRU6516 %IL,FXWSRLQW **77*7$&$$*$*7$7*$$$*$*7$$77$7$77&&*$&7***$&$7&&*&77&& 7**$$*$*&&$$$**&&&7**7$$$$$$$&$&7&77777777&7$$$&$&&7&7& 7&&7*$&77*&&$$777&77&$ Figure 3. $ſ+TGEQIPKVKQPUGSWGPEGCPFEWVRQKPVKPVJGYKNFV[RGNPHS2 gene. The 419delG mutation is noted in red. *HOHOHFWURSKRUHVLV DJDURVH ZDVXVHGYLVXDOL]H Results WKH3&5SURGXFWV*HOVZHUHSKRWRJUDSKHGZKLOH XQGHUXOWUDYLROHWWUDQVOXPLQDWLRQ7KHSUHGLFWHG $WRWDORIVXEMHFWVFRQVLVWLQJRI6516FDVHV ES3&5SURGXFWLVVKRZQLQFigure 2. DQGFRQWUROVXEMHFWVZHUHHQUROOHGGXULQJWKH The NPHS2 GHO*DOOHOHZDVGHWHFWHGE\ VWXG\SHULRG7KHPDMRULW\ZHUHER\VLQ6516JURXS 5)/3DQDO\VLV7KHES3&5SURGXFWVIURPWKH DQGLQWKHFRQWUROJURXS +LVWRU\RI $5063&5DQDO\VLVZHUHVXEMHFWHGWR%IL,UHVWULFWLRQ DWRS\ZDVPRUHFRPPRQLQWKH6516JURXS HQGRQXFOHDVH7KHZLOGW\SHVHTXHQFH ZLWKWKH* WKDQLQWKHFRQWUROJURXS ,QDGGLWLRQWKH SUHVHQW ZDVFXWLQWRWZRIUDJPHQWVRIESDQG NPHS2JHQHPXWDWLRQVZHUHPRUHFRPPRQLQWKH ES7KHPXWDWHGVHTXHQFH *GHOHWHG UHPDLQHG 6516JURXSWKDQLQWKHFRQWUROJURXS(Table 1) XQGLJHVWHGE\%IL,5HVWULFWLRQDQDO\VLVSURGXFWVZHUH 0XOWLYDULDWHDQDO\VHVRIWKHSRVVLEOHULVNIDFWRUV observed by gel electrophoresis as described above. IRU6516DUHGHVFULEHGLQTable 22IWKHVHULVN 7KH%IL,UHFRJQLWLRQVHTXHQFHORFDWHGLQWKHZLOG IDFWRUVPDOHJHQGHUDQGWKHNPHS2&oT and type NPHS2JHQH ES3&5SURGXFW LVVKRZQ GHO*JHQHPXWDWLRQVZHUHVLJQLILFDQWO\DVVRFLDWHG in Figure 3. ZLWKWKHRFFXUUHQFHRI6516 3 +RZHYHU WKHKLVWRU\RIDWRS\ZDVQRWVLJQLILFDQWO\DVVRFLDWHG Table 1. Characteristics of subjects ZLWK6516 3! SNRS group Control group Variable N = 88 N = 65 Gender, n (%) Discussion Female 20 (22.7) 26 (40.0) Male 68 (77.3) 39 (60.0) History of atopy, n (%) 6516LVDFRPPRQFDXVHRIFKURQLFNLGQH\GLVHDVH Positive 25 (28.4) 8 (12.3) LQFKLOGUHQ$FFRUGLQJWRWKHOLWHUDWXUHPXWDWLRQ Negative 63 (71.6) 57 (87.7) in the NPHS2JHQHHQFRGLQJIRUSRGRFLQPD\OHDG 412C T mutation, n (%) o 5,6,7 Positive 58 (65.9) 9 (13.8) to SRNS. $FDVHFRQWUROVWXG\ZDVSHUIRUPHG Negative 30 (34.1) 56 (86.2) LQFKLOGUHQSUHVHQWLQJZLWK6516LQ,QGRQHVLDQ 419delG mutation, n (%) WHDFKLQJKRVSLWDOVIURP6HSWHPEHUWR'HFHPEHU Positive 69 (78.4) 41 (63.1) Negative 19 (21.6) 24 (36.9) 2XUVWXG\UHVXOWVZHUHVLPLODUWRWKRVHIURP Table 2. The variables as risk factors for SRNS both ArabicDQG&DXFDVLDQFKLOGUHQ4,11 The strong UHODWLRQVKLSEHWZHHQWKHRFFXUUHQFHRI6516DQG Variable 14 95% CI P value NPHS2JHQHPXWDWLRQHVSHFLDOO\LQH[RQ Gender 2.21 1.07-4.56 0.036 &oT, was shown by Frishberg et al. in Israeli-Arab 412CoT mutation 18.07 6.76-48.31 < 0.001 419delG mutation 4.55 1.66-12.47 0.003 children, DQGE\&DULGLet alLQ&DXFDVLDQFKLOGUHQ 11 History of atopy 1.81 0.64-5.09 0.262 in Italy. 6LPLODUO\ZHIRXQGWKDWVXEMHFWVZLWKWKH Note: Accuracy 74.51% NPHS2&o7JHQHPXWDWLRQKDGWLPHV 274Paediatr Indones, Vol. 51, No. 5, September 2011 'HGL5DFKPDGLHWDO13+6JHQHPXWDWLRQDWRS\DQGJHQGHUDVULVNIDFWRUVIRU6516 KLJKHUULVNIRU6516WKDQWKRVHZLWKRXWWKLVJHQH ZLWKVSRUDGLF6516WKHPDOHWRIHPDOHUDWLRRI PXWDWLRQ7KHNPHS2&o7JHQHPXWDWLRQ patients with an NPHS2JHQHPXWDWLRQZDV11 LVDQRQVHQVHPXWDWLRQ7KHFKDQJHRIF\WRVLQHWR ,WLVGLIILFXOWWRGLIIHUHQWLDWHEHWZHHQHDUO\)6*6 WK\PLQHDWEDVHVHTXHQFHUHVXOWVLQWKHORVVRI DQGPLQLPDOFKDQJHQHSKURWLFV\QGURPH 0&16 WKHDPLQRDFLGDUJLQLQHDWVHTXHQFH 5; DV)6*6LVFRQVLGHUHGWREHDFRQWLQXDWLRQRI 7KLVDPLQRDFLGORVVSURGXFHVDWUXQFDWHGIRUPRI 0&1617 The relationship between male gender SRGRFLQ,WLVWKRXJKWWKDWWKHWUXQFDWHGSRGLFLQJHWV DQGQHSKURWLFV\QGURPHUHPDLQVXQFOHDU,WKDV WUDSSHGLQWKHHQGRSODVPLFUHWLFXOXPVRWKDWLWORVHV EHHQHVWDEOLVKHGWKDWSDWKRJHQHVLVRIQHSKURWLF LWVFDSDELOLW\WRELQGQHSKULQLQWKHOLSLGUDIW The V\QGURPHLQYROYHV7FHOOG\VIXQFWLRQ Abnormal WUXQFDWHGSRGRFLQZLOOVWLPXODWHDQWLERG\IRUPDWLRQ T cell clones are predominantly located in the DJDLQVWWKHWHUPLQDOSDUWRIWKHSURWHLQ14,15 WK\PXVDQGWK\PXVGLVHDVHLVPRUHFRPPRQLQ :HDOVRFRQVLGHUHGWKHHIIHFWRIDNPHS2 gene boys than girls. Another theory is that SRNS with PXWDWLRQLQDQRWKHUORFDWLRQ8VLQJWKH$5063&5 JHQHPXWDWLRQLVLQIOXHQFHGE\KRUPRQHV(VWURJHQ PHWKRGWKHH[RQ&o7ZDVWKHRQO\PXWDWLRQ PD\SUHYHQWWKHJORPHUXORVFOHURVLVSURFHVVE\ DEOHWREHGHWHFWHG&DULGLet
Load more

Recommended publications
  • Genotyping of Breech Flystrike Resource – Update
    Project No: ON-00515 Contract No: PO4500010753 AWI Project Manager: Bridget Peachey Contractor Name: CSIRO Agriculture and Food Prepared by: Dr Sonja Dominik Publication Date: July 2019 Genotyping of breech flystrike resource – update Published by Australian Wool Innovation Limited, Level 6, 68 Harrington Street, THE ROCKS, NSW, 2000 This publication should only be used as a general aid and is not a substitute for specific advice. To the extent permitted by law, we exclude all liability for loss or damage arising from the use of the information in this publication. AWI invests in research, development, innovation and marketing activities along the global supply chain for Australian wool. AWI is grateful for its funding, which is primarily provided by Australian woolgrowers through a wool levy and by the Australian Government which provides a matching contribution for eligible R&D activities © 2019 Australian Wool Innovation Ltd. All rights reserved. Contents Executive Summary .................................................................................................................... 3 1 Introduction/Hypothesis .................................................................................................... 5 2 Literature Review ............................................................................................................... 6 3 Project Objectives .............................................................................................................. 8 4 Success in Achieving Objectives ........................................................................................
    [Show full text]
  • Evaluation of Variability in Human Kidney Organoids
    ARTICLES https://doi.org/10.1038/s41592-018-0253-2 Evaluation of variability in human kidney organoids Belinda Phipson 1, Pei X. Er1, Alexander N. Combes1,2, Thomas A. Forbes1,3,4, Sara E. Howden1,2, Luke Zappia1,5, Hsan-Jan Yen1, Kynan T. Lawlor1, Lorna J. Hale1,4, Jane Sun6, Ernst Wolvetang6, Minoru Takasato1,7, Alicia Oshlack1,5 and Melissa H. Little 1,2,4* The utility of human pluripotent stem cell–derived kidney organoids relies implicitly on the robustness and transferability of the protocol. Here we analyze the sources of transcriptional variation in a specific kidney organoid protocol. Although individ- ual organoids within a differentiation batch showed strong transcriptional correlation, we noted significant variation between experimental batches, particularly in genes associated with temporal maturation. Single-cell profiling revealed shifts in neph- ron patterning and proportions of component cells. Distinct induced pluripotent stem cell clones showed congruent transcrip- tional programs, with interexperimental and interclonal variation also strongly associated with nephron patterning. Epithelial cells isolated from organoids aligned with total organoids at the same day of differentiation, again implicating relative matura- tion as a confounder. This understanding of experimental variation facilitated an optimized analysis of organoid-based disease modeling, thereby increasing the utility of kidney organoids for personalized medicine and functional genomics. he ability to derive induced pluripotent stem cells (iPSCs) In this study, we provide a comprehensive transcriptional from the somatic cells of patients1, together with directed dif- and morphological evaluation of our kidney organoid protocol. Tferentiation protocols, provides a capacity to model the cell Applying RNA sequencing (RNA-seq) to 57 whole organoids and types affected by disease.
    [Show full text]
  • Nephrin Mutations Can Cause Childhood-Onset Steroid-Resistant Nephrotic Syndrome
    BRIEF COMMUNICATION www.jasn.org Nephrin Mutations Can Cause Childhood-Onset Steroid-Resistant Nephrotic Syndrome Aure´lie Philippe,*† Fabien Nevo,*† Ernie L. Esquivel,*† Dalia Reklaityte,*† ʈ Olivier Gribouval,*† Marie-Jose`phe Teˆte,*‡ Chantal Loirat,§ Jacques Dantal, Michel Fischbach,¶ Claire Pouteil-Noble,** Ste´phane Decramer,†† Martin Hoehne,‡‡ Thomas Benzing,‡‡ Marina Charbit,‡ Patrick Niaudet,*†‡ and Corinne Antignac*†§§ † *Inserm U574, Hoˆpital Necker-Enfants Malades, Universite´ Paris Descartes, Faculte´deMe´ decine Rene´ Descartes, BRIEF COMMUNICATION ‡Pediatric Nephrology and §§Department of Genetics, Hoˆpital Necker-Enfants Malades, Assistance Publique-Hoˆpitaux de Paris, and §Pediatric Nephrology Department, Universite´ Paris VII, Assistance Publique-Hoˆpitaux de Paris, Hoˆpital ʈ Robert Debre´, Paris, ITERT, Department of Nephrology and Clinical Immunology, CHU Nantes, Nantes, ¶Nephrology Dialysis Transplantation Children’s Unit, Hoˆpital de Hautepierre, Strasbourg, **Transplantation and Nephrology Unit, Centre Hospitalier Lyon-Sud, Pierre-Be´nite, and ††Department of Pediatric Nephrology, Hoˆpital des Enfants, and Inserm, U858/I2MR, Department of Renal and Cardiac Remodeling, Toulouse, France; and ‡‡Department of Medicine IV, University of Cologne, Cologne, Germany ABSTRACT Classically, infants with mutations in NPHS1, which encodes nephrin, present with but has since been described in other nephrotic syndrome within the first 3 mo of life (congenital nephrotic syndrome of populations.3–5 Nephrin is a single-pass the Finnish-type), and children with mutations in NPHS2, which encodes podocin, transmembrane protein consisting of present later with steroid-resistant nephrotic syndrome. Recently, however, eight extracellular Ig-like modules, a fi- NPHS2 mutations have been identified in children with congenital nephrotic syn- bronectin type III–like motif, and a cy- drome. Whether NPHS1 mutations similarly account for some cases of childhood tosolic C-terminal tail.
    [Show full text]
  • NPHS2 Antibody (Podocin) (R30382)
    NPHS2 Antibody (Podocin) (R30382) Catalog No. Formulation Size R30382 0.5mg/ml if reconstituted with 0.2ml sterile DI water 100 ug Bulk quote request Availability 1-3 business days Species Reactivity Human, Mouse, Rat Format Antigen affinity purified Clonality Polyclonal (rabbit origin) Isotype Rabbit IgG Purity Antigen affinity Buffer Lyophilized from 1X PBS with 2.5% BSA and 0.025% sodium azide/thimerosal UniProt Q9NP85 Applications Western blot : 0.5-1ug/ml Limitations This NPHS2 antibody is available for research use only. Western blot testing of NPHS2 antibody and rat kidney tissue lysate. Predicted molecular weight ~42kDa. Description NPHS2, also called Podocin (PDCN), is a protein which lines the podocytes and assists in maintaining the barrier at the glomerular basement membrane. NPHS2 is a causative gene for Familial idiopathic nephrotic syndromes, which represents a heterogeneous group of kidney disorders, and include autosomal recessive steroid-resistant nephrotic syndrome, which is characterized by early childhood onset of proteinuria, rapid progression to end-stage renal disease and focal segmental glomerulosclerosis. By positional cloning, it was mapped to 1q25-31. It is almost exclusively expressed in the podocytes of fetal and mature kidney glomeruli, and encodes a new integral membrane protein, podocin, belonging to the stomatin protein family. Boute et al.(2000) found ten different NPHS2 mutations, comprising nonsense, frameshift and missense mutations, to segregate with the disease, demonstrating a crucial role for podocin in the function of the glomerular filtration barrier. Application Notes The stated application concentrations are suggested starting amounts. Titration of the NPHS2 antibody may be required due to differences in protocols and secondary/substrate sensitivity.
    [Show full text]
  • CG/CA Genotypes Represent Novel Markers in the NPHS2 Gene Region Associated with Nephrotic Syndrome
    Journal of Genetics (2020)99:33 Ó Indian Academy of Sciences https://doi.org/10.1007/s12041-020-1188-9 (0123456789().,-volV)(0123456789().,-volV) RESEARCH ARTICLE CG/CA genotypes represent novel markers in the NPHS2 gene region associated with nephrotic syndrome LEILA ESMAELI CHAMGORDANI, NASIM EBRAHIMI, FARZANE AMIRMAHANI and SADEQ VALLIAN* Genetics Division, Faculty of Biological Sciences and Technologies, Department of Cellular and Molecular Biology and Microbiology, University of Isfahan, Isfahan, Iran *For correspondence. E-mail: [email protected]. Received 15 November 2019; revised 10 January 2020; accepted 14 January 2020 Abstract. Nephrotic syndrome (NS) is considered as a primary disease of the kidney that represents a heterogeneous group of glomerular disorders occurring mainly in children. It is generally divided into steroid-sensitive and steroid-resistant forms, depending upon the patient’s response to steroid therapy. Among the genes involved, the NPHS2 gene has been reported as the causative gene in steroid resistant form of nephrotic syndrome. In the present study, heterozygosity rate, allelic frequency and linkage of rs2274625 and rs3829795 markers were investigated in the NPHS2 gene region. To determine the SNP alleles, tetra-primer ARMS PCR was used. After genotyping rs2274625 and rs3829795 polymorphic markers in 120 unrelated individuals and nine trios families, the data were analysed using various computer programs such as UCSC Genome Browser, dbSNP and SNPper. Based on the statistical analysis of the results, for rs2274625 marker, allele frequency for C and T alleles was 97% and 3%, respectively. For rs3829795 marker allele frequency for G and A alleles was 55% and 45%, respectively.
    [Show full text]
  • NPHS2 (Podocin) Mutations in Nephrotic Syndrome
    0031-3998/05/5705-0054R PEDIATRIC RESEARCH Vol. 57, No. 5, Pt 2, 2005 Copyright © 2005 International Pediatric Research Foundation, Inc. Printed in U.S.A. NPHS2 (Podocin) Mutations in Nephrotic Syndrome. Clinical Spectrum and Fine Mechanisms GIANLUCA CARIDI, FRANCESCO PERFUMO, AND GIAN MARCO GHIGGERI Laboratory on Pathophysiology of Uremia [G.C., G.M.C.], and Renal Unit [F.P., G.M.C.], Istituto Giannina Gaslini, 16148 Genova, Italy. ABSTRACT Nephrotic syndrome (NS) is the most frequent cause of iments with the common R229Q polymorphism demonstrated an proteinuria in children and is emerging as a leading cause of altered interaction with nephrin that affects the stability of the uremia. Molecular studies in families with recessive NS have led functional unit. Overall, data are here presented that underscore to the discovery of specialized molecules endowed in podocytes a major role of inherited defects of NPHS2 in NS in children that play a role in proteinuria. This review focalizes the key (including a relevant impact in sporadic cases) and give the position of podocin (NPHS2 gene) in this rapidly evolving field functional rationale for the association. A practical compendium and furnishes a compendium to those involved in clinics and is also given to clinicians involved in the management of NS that genetics of NS. Screening for NPHS2 mutations have been done should modify the classic therapeutic approach. (Pediatr Res 57: in sporadic NS and familial cases with recessive inheritance, 54R–61R, 2005) documenting a mutation detection rate of 45–55% in families and 8–20% in sporadic NS according to the different groups and considering all the clinical phenotypes.
    [Show full text]
  • Genetic Disorders of the Glomerular Filtration Barrier
    CJASN ePress. Published on April 23, 2020 as doi: 10.2215/CJN.11440919 Genetic Disorders of the Glomerular Filtration Barrier Anna S. Li,1,2 Jack F. Ingham,1 and Rachel Lennon 1,3 Abstract The glomerular filtration barrier is a highly specialized capillary wall comprising fenestrated endothelial cells, podocytes, and an intervening basement membrane. In glomerular disease, this barrier loses functional integrity, allowing the passage of macromolecules and cells, and there are associated changes in both cell morphology and the extracellular matrix. Over the past three decades there has been a transformation in our understanding about glomerular disease, fueled by genetic discovery, and this is leading to exciting advances in our knowledge about 1Division of Cell- glomerular biology and pathophysiology. In current clinical practice, a genetic diagnosis already has important Matrix Biology and implications for management ranging from estimating the risk of disease recurrence post-transplant to the life- Regenerative changing advances in the treatment of atypical hemolytic uremic syndrome. Improving our understanding about Medicine, Wellcome Centre for Cell-Matrix the mechanistic basis of glomerular disease is required for more effective and personalized therapy options. In this Research, School of review we describe genotype and phenotype correlations for genetic disorders of the glomerular filtration barrier, Biological Sciences, with a particular emphasis on how these gene defects cluster by both their ontology and patterns of glomerular Faculty of Biology, pathology. Medicine and Health, CJASN ccc–ccc Manchester Academic 15: , 2020. doi: https://doi.org/10.2215/CJN.11440919 Health Science Centre, The University of Manchester, Introduction fenestrae that are 70–100 nm in diameter and covered Manchester, United .
    [Show full text]
  • Steroid-Resistant Nephrotic Syndrome (NPHS2)
    Steroid-Resistant Nephrotic Syndrome (NPHS2) Contact details Introduction Molecular Genetics Service Nephrotic syndrome is a condition that is caused by any of a group of diseases that Level 6, York House damage the kidneys' filtering system, the glomeruli. The structure of the glomeruli 37 Queen Square prevents most protein from getting filtered through into the urine. Nephrotic syndrome London, WC1N 3BH is characterised by proteinuria (abnormally high loss of protein in the urine), T +44 (0) 20 7762 6888 albuminemia and hypercholesterolemia. Ultimately, there is rapid progression to end- F +44 (0) 20 7813 8578 stage renal disease where the kidneys are irreversibly damaged, resulting in death if untreated. Samples required Treatment is based on relieving symptoms, preventing complications and delaying • 5ml venous blood in plastic EDTA progressive kidney damage. Patients with nephrotic syndrome are typically treated bottles (>1ml from neonates) with steroids, of which about 80% have a good response; the rest are considered to • Prenatal testing must be arranged be steroid-resistant and may require renal transplant. One of the main features in in advance, through a Clinical steroid-resistant nephrotic syndrome (SRNS; MIM 600995) is focal segmental Genetics department if possible. glomerulosclerosis (FSGS). • Amniotic fluid or CV samples Mutations in the podocin gene, NPHS2, are associated with autosomal recessive should be sent to Cytogenetics for steroid-resistant nephrotic syndrome (SRNS), including focal segmental dissecting and culturing, with glomerulosclerosis (FSGS). Around half of familial forms and 10-30% of sporadic instructions to forward the sample to the Regional Molecular Genetics forms of SRNS are found to have NPHS2 mutations in both alleles.
    [Show full text]
  • NPHS2 Mutations Account for Only 15% of Nephrotic Syndrome Cases Mara Sanches Guaragna1*, Anna Cristina GB Lutaif2, Vera MS Belangero2, Andréa T
    Guaragna MS, Lutaif ACGB, Belangero VMS, Maciel-Guerra AT, Guerra-Junior G, De Mello MP. Journal of J Rare Dis Res Treat. (2017) 2(3): 49-51 Rare Diseases Research www.rarediseasesjournal.com & Treatment Commentary Open Access Commentary: NPHS2 mutations account for only 15% of nephrotic syndrome cases Mara Sanches Guaragna1*, Anna Cristina GB Lutaif2, Vera MS Belangero2, Andréa T. Maciel-Guerra3,4, Gil Guerra-Junior4,5 and Maricilda P. De Mello1 1Center for Molecular Biology and Genetic Engineering – CBMEG, State University of Campinas, UNICAMP, Campinas, Brazil 2Integrated Center of Pediatric Nephrology – CIN - School of Medical Sciences – FCM, State University of Campinas, UNICAMP, Campinas, Brazil 3Department of Medical Genetics - School of Medical Sciences – FCM, State University of Campinas, UNICAMP, Campinas, Brazil 4Interdisciplinary Group for the Study of Sex Determination and Differentiation – GIEDDS - State University of Campinas, UNICAMP, Campinas, Brazil 5Pediatrics Endocrinology, Department of Pediatrics, School of Medical Sciences – FCM, State University of Campinas, UNICAMP, Campinas, Brazil Article Info Background Article Notes Nephrotic syndrome (NS) represents one of the most common Received: April 30, 2017 kidney conditions that affect children. It manifests as proteinuria, Accepted: May 24, 2017 edema, hypoalbuminemia and hyperlipidemia. According to *Correspondence: the response to standardized corticosteroid therapy, 80 to 90% Caixa Postal 6010 - CBMEG-UNICAMP - 13083-875 - Campinas- SP, Brasil, Tel.: + 55 19 3521 1091; Fax: + 55 19 3521 1089, steroid sensitive (SSNS), whereas the remaining 10 to 20% that are E-mail: [email protected] of children that respond well to the treatment are classified as © 2017 Mara Sanches Guaragna. This article is distributed (SRNS)1.
    [Show full text]
  • Maternal Environment Interacts with Modifier Genes to Influence Progression of Nephrotic Syndrome
    BASIC RESEARCH www.jasn.org Maternal Environment Interacts with Modifier Genes to Influence Progression of Nephrotic Syndrome Julien Ratelade,*† Tiphaine Aguirre Lavin,*† Andrea Onetti Muda,*‡ Ludivine Morisset,*† ʈ ʈ Ge´raldine Mollet,*† Olivia Boyer,*†§ Deborah S. Chen,*† Anna Henger, Matthias Kretzler, Norbert Hubner,¶ Clotilde The´ry,** Marie-Claire Gubler,*† Xavier Montagutelli,†† Corinne Antignac,*†‡‡ and Ernie L. Esquivel*† *INSERM, U574, Hoˆpital Necker-Enfants Malades, †Universite´ Paris Descartes, Faculte´deMe´ decine Rene´ Descartes, Departments of §Pediatric Nephrology and ‡‡Genetics, Hoˆpital Necker-Enfants Malades, Assistance Publique- Hoˆpitaux de Paris, **INSERM, U653, Institut Curie, and ††Unite´deGe´ne´ tique des Mammife`res, Institut Pasteur, ʈ Paris, France; ‡Department of Pathology, Campus Biomedico University, Rome, Italy; Section of Nephrology, University of Michigan School of Medicine, Ann Arbor, Michigan; and ¶Max-Delbruck-Center for Molecular Medicine, Berlin-Buch, Germany ABSTRACT Mutations in the NPHS2 gene, which encodes podocin, are responsible for some cases of sporadic and familial autosomal recessive steroid-resistant nephrotic syndrome. Inter- and intrafamilial variability in the progression of renal disease among patients bearing NPHS2 mutations suggests a potential role for modifier genes. Using a mouse model in which the podocin gene is constitutively inactivated, we sought to identify genetic determinants of the development and progression of renal disease as a result of the nephrotic syndrome. We
    [Show full text]
  • Broadening the Spectrum of Diseases Related to Podocin Mutations
    J Am Soc Nephrol 14: 1278–1286, 2003 Broadening the Spectrum of Diseases Related to Podocin Mutations GIANLUCA CARIDI,* ROBERTA BERTELLI,* MARCO DI DUCA,* MONICA DAGNINO,* FRANCESCO EMMA,‡ ANDREA ONETTI MUDA,§ FRANCESCO SCOLARI,¶ NUNZIA MIGLIETTI,ʈ GIANNA MAZZUCCO,# LUISA MURER,@ ALBA CARREA,* LAURA MASSELLA,‡ GIANFRANCO RIZZONI,‡ FRANCESCO PERFUMO,† and GIAN MARCO GHIGGERI*† *Laboratory on Pathophysiology of Uremia, and †Department of Nephrology, Istituto G. Gaslini, Genova, Italy; ‡Section of Nephrology, Bambin Gesu`Children Hospital, Rome, Italy; §Department of Experimental Medicine and Pathology, University “La Sapienza,” Rome, Italy; ¶Unit of Nephrology, Spedali Civili di Brescia, Italy; ʈDepartment of Pediatrics, University of Brescia, Italy; #Pathology Section, Department of Biomedical Sciences and Human Oncology, University of Torino, Italy; @Department of Pediatrics, University of Padova, Italy. Abstract. A total of 179 children with sporadic nephrotic father, respectively. Immunohistochemistry with anti-podocin syndrome were screened for podocin mutations: 120 with antibodies revealed markedly decreased expression of the pro- steroid resistance, and 59 with steroid dependence/frequent tein in their kidneys. All carriers of heterozygous coding podo- relapses. Fourteen steroid-resistant patients presented homozy- cin mutation or R229Q were screened for nephrin mutation that gous mutations that were associated with early onset of pro- was found in heterozygosity associated with R229Q in one teinuria and variable renal lesions, including one case with patient. Finally, podocin loss of heterozygosity was excluded mesangial C3 deposition. Single mutations of podocin were in one heterozygous child by characterizing cDNA from dis- found in four steroid-resistant and in four steroid-dependent; sected glomeruli. These data outline the clinical features of five patients had the same mutation (P20L).
    [Show full text]
  • Novel Mutations in NPHS2 Detected in Both Familial and Sporadic Steroid-Resistant Nephrotic Syndrome
    J Am Soc Nephrol 13: 388–393, 2002 Novel Mutations in NPHS2 Detected in Both Familial and Sporadic Steroid-Resistant Nephrotic Syndrome STEPHANIE M. KARLE, BARBARA UETZ, VERA RONNER, LISA GLAESER, FRIEDHELM HILDEBRANDT, ARNO FUCHSHUBER, and the Arbeitsgemeinschaft fu¨r Pa¨diatrische Nephrologie Study Group University Children’s Hospital, Freiburg, Germany. Abstract. Autosomal recessive steroid-resistant nephrotic syn- R196P, V290M, IVS4-1G3T, and 460-467insT in 12 (46%) drome (SRINS) belongs to the heterogeneous group of familial of 26 multiplex families and in 7 (28%) of 25 single patients nephrotic syndrome and represents a frequent cause of end- with the clinical diagnosis of a SRINS. Because NPHS2 mu- stage renal disease in childhood. This kidney disorder is char- tations were found in nearly 30% of these patients with “spo- acterized by early onset of proteinuria, progression to end- radic” SRINS, mutational analysis should also be performed in stage renal disease, and histologic findings of focal segmental these patients. Besides better classification of the disease en- glomerulosclerosis, minimal change nephrotic syndrome, or tity, identification of NPHS2 mutations may save some of these both. A causative gene, NPHS2, has been mapped to chromo- patients from unnecessary steroid treatment and also permit the some 1q25-q31 and was recently identified by positional clon- prediction of absence of disease recurrence after kidney ing. This study reports five novel NPHS2 mutations: A284V, transplantation. Autosomal recessive steroid-resistant nephrotic syndrome and single patients with SRINS to determine whether NPHS2 (SRINS) belongs to the heterogeneous group of familial ne- mutations are also a cause of sporadic SRINS.
    [Show full text]