title: Very early onset inflammatory bowel disease associated with aberrant trafficking of IL-10R1 and cure by T cell replete haploidentical bone marrow transplantation.
abstract: OBJECTIVE
  Loss-of-function mutations in IL10 and IL10R cause very early onset inflammatory bowel disease (VEO-IBD). Here, we investigated the molecular pathomechanism of a novel intronic IL10RA mutation and describe a new therapeutic approach of T cell replete haploidentical hematopoietic stem cell transplantation (HSCT).
  
  
  METHODS
  Clinical data were collected by chart review. Genotypes of IL10 and IL10R genes were determined by Sanger sequencing. Expression and function of mutated IL-10R1 were assessed by quantitative PCR, Western blot analysis, enzyme-linked immunosorbent assays, confocal microscopy, and flow cytometry.
  
  
  RESULTS
  We identified a novel homozygous point mutation in intron 3 of the IL10RA (c.368-10C > G) in three related children with VEO-IBD. Bioinformatical analysis predicted an additional 3' splice site created by the mutation. Quantitative PCR analysis showed normal mRNA expression of mutated IL10RA. Sequencing of the patient's cDNA revealed an insertion of the last nine nucleotides of intron 3 as a result of aberrant splicing. Structure-based modeling suggested misfolding of mutated IL-10R1. Western blot analysis demonstrated a different N-linked glycosylation pattern of mutated protein. Immunofluorescence and FACS analysis revealed impaired expression of mutated IL-10R1 at the plasma membrane. In the absence of HLA-identical donors, T cell replete haploidentical HSCT was successfully performed in two patients.
  
  
  CONCLUSIONS
  Our findings expand the spectrum of IL10R mutations in VEO-IBD and emphasize the need for genetic diagnosis of mutations in conserved non-coding sequences of candidate genes. Transplantation of haploidentical stem cells represents a curative therapy in IL-10R-deficient patients, but may be complicated by non-engraftment.
authors: Dr. von Hauner Children's Hospital, Ludwig Maximilians University, Munich, Germany.
authors: Dhaarini
authors: D
authors: Murugan
authors: Michael H
authors: MH
authors: Albert
authors: Jörg
authors: J
authors: Langemeier
authors: Jens
authors: J
authors: Bohne
authors: Jacek
authors: J
authors: Puchalka
authors: Päivi M
authors: PM
authors: Järvinen
authors: Fabian
authors: F
authors: Hauck
authors: Anne K
authors: AK
authors: Klenk
authors: Christine
authors: C
authors: Prell
authors: Stephanie
authors: S
authors: Schatz
authors: Jana
authors: J
authors: Diestelhorst
authors: Barbara
authors: B
authors: Sciskala
authors: Naschla
authors: N
authors: Kohistani
authors: Bernd H
authors: BH
authors: Belohradsky
authors: Susanna
authors: S
authors: Müller
authors: Thomas
authors: T
authors: Kirchner
authors: Mark R
authors: MR
authors: Walter
authors: Philip
authors: P
authors: Bufler
authors: Aleixo M
authors: AM
authors: Muise
authors: Scott B
authors: SB
authors: Snapper
authors: Sibylle
authors: S
authors: Koletzko
authors: Christoph
authors: C
authors: Klein
authors: Daniel
authors: D
authors: Kotlarz
mesh_terms: D017668:Age of Onset; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D016026:Bone Marrow Transplantation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D003241:Consanguinity; D004252:DNA Mutational Analysis; D005260:Female; D005838:Genotype; D006031:Glycosylation; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000628:therapy*; D053709:Interleukin-10 Receptor alpha Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007438:Introns; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D010641:Phenotype; D011487:Protein Conformation; D021381:Protein Transport; D016415:Sequence Alignment; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology; D016896:Treatment Outcome
chemical_list: D053709:Interleukin-10 Receptor alpha Subunit