GENOTYPE–PHENOTYPE CORRELATIONS IN NONLETHAL OSTEOGENESIS IMPERFECTA CAUSED BY MUTATIONS IN THE HELICAL DOMAIN OF COLLAGEN TYPE I Frank Rauch, Liljana Lalic, Peter Roughley and Francis H Glorieux Nur Rohmah 12/338710/PMU/7375

Introduction heritable connective tissue disorder Osteogenesis Imperfecta heritable connective tissue disorder mutation in COL1A1 or COL1A2

Introduction

Introduction Hearing loss Osteogenesis Imperfecta (OI) Blue/grey sclera bone fragility Dentinogenesis imperfecta

Introduction Type I < IV < III < II Type Clinical severity Typical features I Mild non-deforming OI Normal height or mild short stature; blue sclera; no dentinogenesis imperfecta II Perinatal lethal Multiple rib and long-bone fractures at birth; pronounced deformities; broad long bones, low density of skull bones on radiographs; dark sclera III Severely deforming Very short; triangular face; severe scoliosis; greyish sclera; dentinogenesis imperfecta IV Moderately deforming Moderately short; mild to moderate scoliosis; greyish or white sclera; dentinogenesis imperfecta

Objective to investigated genotype–phenotype correlations in OI patients with glycine mutations in the triple helical domain of collagen type I.

Subject and method Sample Patient Clinical characteristics Total genomic DNA was isolated from peripheral blood Amplified by PCR and sequencing aligned with the GenBank reference sequences Statistical analysis

Collagen type I mutation analysis Subject and method Sampel Patient 161 OI patients DNA isolation Total genomic DNA was isolated from peripheral blood using the QIAamp DNA Blood Midi Kit Collagen type I mutation analysis All exons of the COL1A1 and COL1A2 genes amplified by PCR. The sequencing reactionusing a BigDye Terminator cycle sequencing kit . The nucleotide sequence was determined using an Applied Biosystems 3100 DNA sequencer.

Subject and method Alignment Sequence traces were aligned with the GenBank reference sequencesCOL1A1 genomic DNA (AF017178) COL1A2 genomic DNA (AF004877.1), then compared and analyzed using BLAST. Statistical analysis

RESULTS AND DISCUSSION

RESULTS AND DISCUSSION

RESULTS AND DISCUSSION

RESULTS AND DISCUSSION Figure 1 Relationship between the triple helical position of glycine mutations in collagen type I a chains and the presence (+) or absence () of deformities or fractures at birth (DFB), blue sclera (BS), and dentinogenesis imperfecta (DI).

RESULTS AND DISCUSSION Figure 2. Relationship between the positions of glycine substitutions in the triple helical domain of type I collagen and height z-scores

Conclusion genotype–phenotype correlations for 161 OI patients with 111 distinct triple helical mutations of collagen type I Height correlated with the location of the mutation in the α2 chain but not in the α1. Patients with mutations affecting the first 120 amino acids at the amino-terminal end of the collagen type I triple helix had blue sclera but did not have dentinogenesis imperfecta.

Dimana: Z = Nilai standar X = Rata-rata distribusi Rumus Z-score : Manual Rumus Z-score : Z= X-M SD Dimana: Z = Nilai standar X = Rata-rata distribusi M = Nilai mentah yang akan dicari nilai standarnya SD = Standar deviasi distribusi