Unique X-linked familial FSGS with co-segregating heart block disorder is associated with a mutation in the NXF5 gene

Teresa Esposito1^, Rod A Lea2^, Bridget H Maher2, Dianne Moses2, Hannah C Cox2, Sara Magliocca1, Andrea Angius3, Dale R Nyholt4, Thomas Titus5, Troy Kay5, Nicholas A Gray6, Maria P Rastaldi7, Alan Parnham5, Fernando Gianfrancesco1+, Lyn R Griffiths2+*

^ Joint First Authors

+ Joint Senior Authors

1Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", National Research Council of Italy, Naples, Italy.

2Genomics Research Centre, Griffith Health Institute, Griffith University, Gold Coast, Queensland, 4222, Australia.

3Institute of Genetic and Biomedical Research, National Research Council of Italy, Cagliari, Italy.

4Queensland Institute of Medical Research, Herston Road, Herston, Brisbane, 4006, Australia

5Renal Department, Gold Coast Hospital, Queensland, Australia

6Renal Services, Nambour Hospital, Nambour, Queensland, Australia

7Renal Research Laboratory, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy

Corresponding author:

*Professor Lyn Griffiths, Genomics Research Centre, Griffith Health Institute, Griffith University, Queensland 4222, Australia.; Phone +61 7 5552 8664, fax +61 7 5552 9081, e-mail:

Supplementary Table. 1 Summary of the molecular genetics of FSGS and progressive heart block

Disorder / Inheritance / Locus / Gene / Protein
FSGS1 / Aut. Dom / 19q13 / ACTN4 / a-actinin 4
FSGS2 / Aut. Dom / 11q21-q22 / TRPC6 / Transient recept. pot. cat. chan. 6
FSGS3 / Aut. Rec / 6p12 / CD2AP / CD2 – associated protein
FSGS4 / Aut. Dom / 22q12 / MYH9 / Myosin, heavy chain 9
NPHS1 / Aut. Rec / 19q13.1 / NPHS1 / Nephrin
NPHS2 / Aut. Rec / 1q25-q31 / NPHS2 / Podocin
NPHS3 / Aut. Rec / 10q23.32-q24.1 / PLCE1 / Phospholipase C Epsilon
FSGS / Aut. Dom / 14q32.33 / INF2 / Formin
FSGS / Aut. Rec / 15q21 / MYO1E / Myosin 1E
PFHB1A / Aut. Dom / 3p21 / SCN5A / Sodium Channel, voltage-gated, type V
PFHB1B / Aut. Dom / 19q13.2-q13.3 / TRPM4 / Transient receptor potential cation channel
PFHB2 / Aut. Dom / 1q32.2-q32.3 / Unknown

Supplementary Table 2. Table of pathologies in the pedigree

Pathology / Individuals
FSGS
Premature death in generations I,II,III – family history suggests renal failure / ID1, ID8, ID12
FSGS not confirmed by renal biopsy* / ID14, ID16, ID50, ID62
FSGS confirmed by renal biopsy / ID69, ID75, ID85, D136
Kidney Transplant / ID62, ID69, ID105
Cardiac Conduction Defects
Cardiac conduction defects / ID14, ID16, ID50, ID69, ID75, ID85
Pacemaker / ID14, ID50, ID69, ID75, ID85
Other
Renal Tumours / ID30, ID34, ID39, ID100
Pre-eclampsia and hypertension / ID34, ID47, ID55, ID59, ID80 ID82, ID84, ID88, ID139, ID142
Asymptomatic proteinuria (carriers) / ID34, ID47, ID55, ID78

* A histological diagnosis of the renal pathology is unavailable for these four affected males as they died prior to the development of renal biopsy as a routine clinical investigation in the regions they were residing.

Supplementary Table. 3 Summary of the molecular genetics analysis of candidate genes located in linkage interval.

Gene Disorder / Protein Inheritance / Disorder / Candidate gene status
GLA / Galactosidase, ALPHA / Fabry Disease / No mutations
CLDN2 / Claudin 2 / No mutations
COL4A5 / Collagen, type IV, ALPHA-5 / Alport Syndrome / No mutations
TRPC5 / Transient Receptor Potential Cation
Channel, Subfamily C, Member 5 / No mutations
AMOT / Angiomotin / No mutations

These five genes were analysed by direct sequencing of all exons including exon-intron boundary. Primers sequences are available on request.

Supplementary Figure 1 The complete 6-generational Australian FSGS pedigree.

Supplementary Figure 2 Genomic map of the candidate interval showing all genes housing this region. Underlined are indicated the candidate genes analysed by direct sequencing.


Supplementary Figure 3: Exome sequencing analysis. A, coverage of part of the COL4A5 gene located in the linkage interval. B sequence of the exon 12 of the COL4A5 gene containing the codon 222 that was mutated in a patient with glomerulopathy.

Supplementary Figure 4 SIFT prediction of the R113W mutation in NXF5 gene. From amino acid 101 to 121 are reported and the position 113 is indicated with a rectangle. At left are indicated the deleterious changes and at right are indicated the tolerated changes.