43 results on '"Sangermano, R"'
Search Results
2. ABCA4-associated disease as a model for missing heritability in autosomal recessive disorders: novel noncoding splice, cis-regulatory, structural, and recurrent hypomorphic variants
- Author
-
Bauwens, Miriam, Garanto, A., Sangermano, R., Naessens, Sarah, Weisschuh, Nicole, Zaeytijd, Julie De, Khan, M., Collin, R.W.J., Cremers, F.P.M., Leroy, Bart P., Baere, Elfride De, Bauwens, Miriam, Garanto, A., Sangermano, R., Naessens, Sarah, Weisschuh, Nicole, Zaeytijd, Julie De, Khan, M., Collin, R.W.J., Cremers, F.P.M., Leroy, Bart P., and Baere, Elfride De
- Abstract
Contains fulltext : 206085.pdf (publisher's version ) (Open Access)
- Published
- 2019
3. Late-Onset Stargardt Disease Due to Mild, Deep-Intronic ABCA4 Alleles
- Author
-
Runhart, E.H., Valkenburg, D., Cornelis, S.S., Khan, M., Sangermano, R., Albert, S., Bax, N.M., Astuti, G.D.N, Gilissen, C.F., Blokland, E.A.W., Cremers, F.P.M., Ingeborgh van den Born, L., Hoyng, C.B., Runhart, E.H., Valkenburg, D., Cornelis, S.S., Khan, M., Sangermano, R., Albert, S., Bax, N.M., Astuti, G.D.N, Gilissen, C.F., Blokland, E.A.W., Cremers, F.P.M., Ingeborgh van den Born, L., and Hoyng, C.B.
- Abstract
Contains fulltext : 209464.pdf (publisher's version ) (Open Access)
- Published
- 2019
4. Deep-intronic ABCA4 variants explain missing heritability in Stargardt disease and allow correction of splice defects by antisense oligonucleotides
- Author
-
Sangermano, R, Garanto, A., Khan, M. (Mubeen), Runhart, E.H., Bauwens, M., Bax, N.M.A. (Klaas), Born, L.I. (Ingeborgh) van den, Khan, M.I. (Muhammad), Cornelis, S.S., Verheij, J, Pott, J.W.R., Thiadens, A., Klaver, C.C.W. (Caroline), Puech, B., Meunier, I., Naessens, S., Arno, G., Fakin, A., Carss, K.J., Raymond, FL, Webster, A.R. (Andrew), Dhaenens, C.M., Stohr, H., Grassmann, F. (Felix), Weber, B.H.F. (Bernhard), Hoyng, C.B. (Carel), De Baere, E. (Elfride), Albert, S., Collin, R.W.J. (Rob), Cremers, F.P.M. (Frans), Sangermano, R, Garanto, A., Khan, M. (Mubeen), Runhart, E.H., Bauwens, M., Bax, N.M.A. (Klaas), Born, L.I. (Ingeborgh) van den, Khan, M.I. (Muhammad), Cornelis, S.S., Verheij, J, Pott, J.W.R., Thiadens, A., Klaver, C.C.W. (Caroline), Puech, B., Meunier, I., Naessens, S., Arno, G., Fakin, A., Carss, K.J., Raymond, FL, Webster, A.R. (Andrew), Dhaenens, C.M., Stohr, H., Grassmann, F. (Felix), Weber, B.H.F. (Bernhard), Hoyng, C.B. (Carel), De Baere, E. (Elfride), Albert, S., Collin, R.W.J. (Rob), and Cremers, F.P.M. (Frans)
- Abstract
Purpose: Using exome sequencing, the underlying variants in many persons with autosomal recessive diseases remain undetected. We explored autosomal recessive Stargardt disease (STGD1) as a model to identify the missing heritability. Methods: Sequencing of ABCA4 was performed in 8 STGD1 cases with one variant and p.Asn1868Ile in trans, 25 cases with one variant, and 3 cases with no ABCA4 variant. The effect of intronic variants was analyzed using in vitro splice assays in HEK293T cells and patient-derived fibroblasts. Antisense oligonucleotides were used to correct splice defects. Results: In 24 of the probands (67%), one known and five novel deep-intronic variants were found. The five novel variants resulted in messenger RNA pseudoexon inclusions, due to strengthening of cryptic splice sites or by disrupting a splicing silencer motif. Variant c.769-784C>T showed partial insertion of a pseudoexon and was found in cis with c.5603A>T (p.Asn1868Ile), so its causal role could not be fully established. Variant c.4253+43G>A resulted in partial skipping of exon 28. Remarkably, antisense oligonucleotides targeting the aberrant splice processes resulted in (partial) correction of all splicing defects. Conclusion: Our data demonstrate the importance of assessing noncoding variants in genetic diseases, and show the great potential of splice modulation therapy for deep-intronic variants.
- Published
- 2019
- Full Text
- View/download PDF
5. Unraveling the missing heritability in ABCA4-associated Stargardt disease
- Author
-
Sangermano, R., Cremers, F.P.M., Albert, S., Collin, R.W.J., and Radboud University Nijmegen
- Subjects
Radboud Institute for Molecular Life Sciences ,Sensory disorders Radboud Institute for Molecular Life Sciences [Radboudumc 12] ,Sensory disorders [Radboudumc 12] - Abstract
Contains fulltext : 191613.pdf (Publisher’s version ) (Open Access) Radboud University, 28 juni 2018 Promotor : Cremers, F.P.M. Co-promotores : Albert, S., Collin, R.W.J.
- Published
- 2018
6. Identification and Rescue of Splice Defects Caused by Two Neighboring Deep-Intronic ABCA4 Mutations Underlying Stargardt Disease
- Author
-
Albert, S., Garanto, A., Sangermano, R., Khan, M., Bax, N.M., Hoyng, C.B., Collin, R.W.J., Cremers, F.P.M., Albert, S., Garanto, A., Sangermano, R., Khan, M., Bax, N.M., Hoyng, C.B., Collin, R.W.J., and Cremers, F.P.M.
- Abstract
Item does not contain fulltext
- Published
- 2018
7. The Common ABCA4 Variant p.Asn1868Ile Shows Nonpenetrance and Variable Expression of Stargardt Disease When Present in trans With Severe Variants
- Author
-
Runhart, E.H., Sangermano, R., Cornelis, S.S., Verheij, J.B.G.M., Plomp, A.S., Boon, C.J.F., Lugtenberg, D., Roosing, S., Bax, N.M., Blokland, E.A.W., Jacobs-Camps, M.H.M., Velde-Visser, S.D. van der, Klaver, C.C.W., Hoyng, C.B., Cremers, F.P.M., Runhart, E.H., Sangermano, R., Cornelis, S.S., Verheij, J.B.G.M., Plomp, A.S., Boon, C.J.F., Lugtenberg, D., Roosing, S., Bax, N.M., Blokland, E.A.W., Jacobs-Camps, M.H.M., Velde-Visser, S.D. van der, Klaver, C.C.W., Hoyng, C.B., and Cremers, F.P.M.
- Abstract
Contains fulltext : 194515.pdf (Publisher’s version ) (Open Access)
- Published
- 2018
8. Unraveling the missing heritability in ABCA4-associated Stargardt disease
- Author
-
Cremers, F.P.M., Albert, S., Collin, R.W.J., Sangermano, R., Cremers, F.P.M., Albert, S., Collin, R.W.J., and Sangermano, R.
- Abstract
Radboud University, 28 juni 2018, Promotor : Cremers, F.P.M. Co-promotores : Albert, S., Collin, R.W.J., Contains fulltext : 191613.pdf (publisher's version ) (Open Access)
- Published
- 2018
9. ABCA4 midigenes reveal the full splice spectrum of all reported noncanonical splice site variants in Stargardt disease
- Author
-
Sangermano, R., Khan, M., Cornelis, S.S., Richelle, Valerie, Albert, S., Garanto, A., Lugtenberg, D., Collin, R.W.J., Cremers, F.P.M., Sangermano, R., Khan, M., Cornelis, S.S., Richelle, Valerie, Albert, S., Garanto, A., Lugtenberg, D., Collin, R.W.J., and Cremers, F.P.M.
- Abstract
Contains fulltext : 183701.pdf (publisher's version ) (Open Access)
- Published
- 2018
10. The Common ABCA4 Variant p.Asn1868Ile Shows Nonpenetrance and Variable Expression of Stargardt Disease When Present in trans With Severe Variants
- Author
-
Runhart, EH, Sangermano, R, Cornelis, SS, Verheij, J, Plomp, AS, Boon, CJF, Lugtenberg, D, Roosing, S, Bax, NMA, Blokland, EAW, Jacobs-Camps, MHM, van der Velde-Visser, SD, Pott, JWR, Rohrschneider, K, Thiadens, Alberta, Klaver, Caroline, van den Born, LI, Hoyng, CB, Cremers, FPM, Runhart, EH, Sangermano, R, Cornelis, SS, Verheij, J, Plomp, AS, Boon, CJF, Lugtenberg, D, Roosing, S, Bax, NMA, Blokland, EAW, Jacobs-Camps, MHM, van der Velde-Visser, SD, Pott, JWR, Rohrschneider, K, Thiadens, Alberta, Klaver, Caroline, van den Born, LI, Hoyng, CB, and Cremers, FPM
- Published
- 2018
11. The treatment of black stain associated with of iron metabolism disorders with lactoferrin: a litterature search and two case studies.
- Author
-
Sangermano, R., Pernarella, S., Straker, M., Lepanto, M. S., Rosa, L., Cutone, A., Valenti, P., and Ottolenghi, L.
- Subjects
IRON metabolism disorders ,LACTOFERRIN ,CHROMOGENIC bacteria ,ORAL diseases ,METABOLIC disorders - Abstract
Among the various pathologies of the oral cavity, the formation of "unsightly black spots" on the surface of the tooth, universally known as Black Stain (BS) has recently been acquiring more interest. Usually BS is typically found in individuals in prepubertal age, even though it has been identified in adults associated with microbial exchange and / or with iron metabolism disorders. Microbial exchange concerns the possible exchange of bacteria between family members which can take place directly, through effusions, or indirectly, through brushes, cutlery or glasses. For this reason, it is recommended that toothbrushes of family members not be left damp and in contact with each other. The bathroom, being a warm-humid environment, is in fact an optimal habitat for microbial proliferation. Of specific importance in BS is the accumulation of iron in tissues and secretions which, together with chromogenic bacteria, are the primary cause of this pathology. In fact, among the metabolic products synthesized by bacteria in the oral cavity, hydrogen sulfide is of considerable interest, since upon reacting with iron available in saliva, in pathological conditions (iron metabolism disorders), it forms black precipitates consisting of ferric sulfide. These precipitates bind to the surface of the teeth, tending to form a stria that usually follows the contour of the gingiva, with an unsightly and variable chromatic intensity. In physiological situations, iron homeostasis is defined as the state of equilibrium between iron present in tissues and in secretions and that which is present in the circulation. Instead, in pathological conditions, defined as iron metabolism disorders, there is an accumulation of iron in tissues and secretions and a lack of it in the circulation. It is also important to remember that subjects affected by BS are more protected from carious processes than healthy subjects, probably due to a significant predominance of chromogenic bacteria compared to those responsible for caries. It should also be remembered that in young subjects BS tends to regress with pubertal development and the transition to adult life. In any case, using common professional hygiene procedures, it is possible to remove BS as well as plaque and tartar deposits. In particular, with ultrasonic scalers, polishing pastes and powders carried by air and water jets, the surfaces of the teeth can be restored to their natural healthy state. All the techniques for removing the precipitates, are not enough however, to fix and permanently eradicate their appearance, as these precipitates last only for short periods and recur very frequently. Due to the frequent recurrences, new oral microbiota control therapies are emerging; among these the use of lactoferrin (Lf) in the dental field and particularly in the treatment of BS appears to be very promising. Taken togheter, here the effect of Lf in subjects affected by BS has been investigated. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
12. Photoreceptor Progenitor mRNA Analysis Reveals Exon Skipping Resulting from the ABCA4 c.5461-10T-->C Mutation in Stargardt Disease
- Author
-
Sangermano, R., Bax, N.M., Bauwens, M., Born, L.I. van den, Baere, E. De, Garanto, A., Collin, R.W.J., Goercharn-Ramlal, A.S.A., Engelsman-van Dijk, A.H.A. den, Rohrschneider, K., Hoyng, C.B., Cremers, F.P.M., Albert, S., Sangermano, R., Bax, N.M., Bauwens, M., Born, L.I. van den, Baere, E. De, Garanto, A., Collin, R.W.J., Goercharn-Ramlal, A.S.A., Engelsman-van Dijk, A.H.A. den, Rohrschneider, K., Hoyng, C.B., Cremers, F.P.M., and Albert, S.
- Abstract
Item does not contain fulltext, PURPOSE: To elucidate the functional effect of the ABCA4 variant c.5461-10T-->C, one of the most frequent variants associated with Stargardt disease (STGD1). DESIGN: Case series. PARTICIPANTS: Seventeen persons with STGD1 carrying ABCA4 variants and 1 control participant. METHODS: Haplotype analysis of 4 homozygotes and 11 heterozygotes for c.5461-10T-->C and sequence analysis of the ABCA4 gene for a homozygous proband. Fibroblasts were reprogrammed from 3 persons with STGD1 into induced pluripotent stem cells, which were differentiated into photoreceptor progenitor cells (PPCs). The effect of the c.5461-10T-->C variant on RNA splicing by reverse-transcription polymerase chain reaction was analyzed using PPC mRNA. In vitro assays were performed with minigene constructs containing ABCA4 exon 39. We analyzed the natural history and ophthalmologic characteristics of 4 persons homozygous for c.5461-10T-->C. MAIN OUTCOME MEASURES: Haplotype and rare variant data for ABCA4, RNA splice defects, age at diagnosis, visual acuity, fundus appearance, visual field, electroretinography (ERG) results, fluorescein angiography results, and fundus autofluorescence findings. RESULTS: The frequent ABCA4 variant c.5461-10T-->C has a subtle effect on splicing based on prediction programs. A founder haplotype containing c.5461-10T-->C was found to span approximately 96 kb of ABCA4 and did not contain other rare sequence variants. Patient-derived PPCs showed skipping of exon 39 or exons 39 and 40 in the mRNA. HEK293T cell transduction with minigenes carrying exon 39 showed that the splice defects were the result of the c.5461-10T-->C variant. All 4 subjects carrying the c.5461-10T-->C variant in a homozygous state showed a young age of STGD1 onset, with low visual acuity at presentation and abnormal cone ERG results. All 4 demonstrated severe cone-rod dystrophy before 20 years of age and were legally blind by 25 years of age. CONCLUSIONS: The ABCA4 variant c.5461-10T-->C is located on a fo
- Published
- 2016
13. Mutations in MFSD8, encoding a lysosomal membrane protein, are associated with nonsyndromic autosomal recessive macular dystrophy
- Author
-
Roosing, S., Born, L.I. van den, Sangermano, R., Banfi, S., Koenekoop, R.K., Zonneveld-Vrieling, M.N., Klaver, C.C., Lith-Verhoeven, J.J. van, Cremers, F.P.M., Hollander, A.I. den, Hoyng, C.B., Roosing, S., Born, L.I. van den, Sangermano, R., Banfi, S., Koenekoop, R.K., Zonneveld-Vrieling, M.N., Klaver, C.C., Lith-Verhoeven, J.J. van, Cremers, F.P.M., Hollander, A.I. den, and Hoyng, C.B.
- Abstract
Item does not contain fulltext, PURPOSE: This study aimed to identify the genetic defects in 2 families with autosomal recessive macular dystrophy with central cone involvement. DESIGN: Case series. PARTICIPANTS: Two families and a cohort of 244 individuals with various inherited maculopathies and cone disorders. METHODS: Genome-wide linkage analysis and exome sequencing were performed in 1 large family with 5 affected individuals. In addition, exome sequencing was performed in the proband of a second family. Subsequent analysis of the identified mutations in 244 patients was performed by Sanger sequencing or restriction enzyme digestion. The medical history of individuals carrying the MFSD8 variants was reviewed and additional ophthalmic examinations were performed, including electroretinography (ERG), multifocal ERG (mfERG), perimetry, optical coherence tomography (OCT), fundus autofluorescence, and fundus photography. MAIN OUTCOME MEASURES: MFSD8 variants, age at diagnosis, visual acuity, fundus appearance, color vision defects, visual field, ERG, mfERG, fundus autofluorescence, and OCT findings. RESULTS: Compound heterozygous variants in MFSD8, a gene encoding a lysosomal transmembrane protein, were identified in 2 families with macular dystrophy with a normal or subnormal ERG, but reduced mfERG. In both families, a heterozygous missense variant p.Glu336Gln was identified, which was predicted to have a mild effect on the protein. In the first family, a protein-truncating variant (p.Glu381*) was identified on the other allele, and in the second family, a variant (c.1102G>C) was identified that results in a splicing defect leading to skipping of exon 11 (p.Lys333Lysfs*3). The p.Glu336Gln allele was found to be significantly enriched in patients with maculopathies and cone disorders (6/488) compared with ethnically matched controls (35/18 682; P < 0.0001), suggesting that it may act as a genetic modifier. CONCLUSIONS: In this study, we identified variants in MFSD8 as a novel cause of nonsyndromic
- Published
- 2015
14. Heterozygous deep-intronic variants and deletions in ABCA4 in persons with retinal dystrophies and one exonic ABCA4 variant
- Author
-
Bax, N.M., Sangermano, R., Roosing, S., Thiadens, A.A.H.J., Hoefsloot, L.H., Born, L.I. van den, Phan, M., Klevering, B.J., Westeneng-van Haaften, S.C., Braun, T.A., Zonneveld-Vrieling, M.N., Wijs, I. de, Mutlu, M., Stone, E.M., Hollander, A.I. den, Klaver, C.C., Hoyng, C.B., Cremers, F.P.M., Bax, N.M., Sangermano, R., Roosing, S., Thiadens, A.A.H.J., Hoefsloot, L.H., Born, L.I. van den, Phan, M., Klevering, B.J., Westeneng-van Haaften, S.C., Braun, T.A., Zonneveld-Vrieling, M.N., Wijs, I. de, Mutlu, M., Stone, E.M., Hollander, A.I. den, Klaver, C.C., Hoyng, C.B., and Cremers, F.P.M.
- Abstract
Item does not contain fulltext, Variants in ABCA4 are responsible for autosomal-recessive Stargardt disease and cone-rod dystrophy. Sequence analysis of ABCA4 exons previously revealed one causative variant in each of 45 probands. To identify the "missing" variants in these cases, we performed multiplex ligation-dependent probe amplification-based deletion scanning of ABCA4. In addition, we sequenced the promoter region, fragments containing five deep-intronic splice variants, and 15 deep-intronic regions containing weak splice sites. Heterozygous deletions spanning ABCA4 exon 5 or exons 20-22 were found in two probands, heterozygous deep-intronic variants were identified in six probands, and a deep-intronic variant was found together with an exon 20-22 deletion in one proband. Based on ophthalmologic findings and characteristics of the identified exonic variants present in trans, the deep-intronic variants V1 and V4 were predicted to be relatively mild and severe, respectively. These findings are important for proper genetic counseling and for the development of variant-specific therapies.
- Published
- 2015
15. Autosomal dominant Ménétrier-like disease.
- Author
-
Strisciuglio C, Corleto VD, Brunetti-Pierri N, Piccolo P, Sangermano R, Rindi G, Martini M, D'Armiento FP, Staiano A, Miele E, Strisciuglio, Caterina, Corleto, Vito D, Brunetti-Pierri, Nicola, Piccolo, Pasquale, Sangermano, Riccardo, Rindi, Guido, Martini, Maurizio, D'Armiento, Francesco P, Staiano, Annamaria, and Miele, Erasmo
- Published
- 2012
- Full Text
- View/download PDF
16. Gene decay: analytic simulation of gene decay
- Author
-
Rossi, C and Sangermano, R
- Subjects
Settore MED/01 - Statistica Medica - Published
- 1972
17. Mutations in MFSD8, encoding a lysosomal membrane protein, are associated with nonsyndromic autosomal recessive macular dystrophy
- Author
-
Carel B. Hoyng, Frans P.M. Cremers, L. Ingeborgh van den Born, Sandro Banfi, Riccardo Sangermano, Susanne Roosing, Marijke N. Zonneveld-Vrieling, Caroline C W Klaver, Janneke J.C. van Lith-Verhoeven, Anneke I. den Hollander, Robert K. Koenekoop, Ophthalmology, Roosing, S, van den Born, Li, Sangermano, R, Banfi, Sandro, Koenekoop, Rk, Zonneveld Vrieling, Mn, Klaver, Cc, van Lith Verhoeven, Jj, Cremers, Fp, den Hollander, Ai, and Hoyng, Cb
- Subjects
Adult ,Male ,Proband ,Pathology ,medicine.medical_specialty ,Achromatopsia ,genetic structures ,DNA Mutational Analysis ,Sensory disorders Radboud Institute for Health Sciences [Radboudumc 12] ,Genome-wide association study ,Compound heterozygosity ,Macular Degeneration ,Cone dystrophy ,Electroretinography ,medicine ,Humans ,Exome ,Sensory disorders Radboud Institute for Molecular Life Sciences [Radboudumc 12] ,Exome sequencing ,Aged ,Genetics ,business.industry ,Lysosome-Associated Membrane Glycoproteins ,Membrane Transport Proteins ,Middle Aged ,Macular dystrophy ,medicine.disease ,eye diseases ,Pedigree ,Ophthalmology ,Mutation ,Visual Field Tests ,Female ,Neuronal ceroid lipofuscinosis ,sense organs ,business ,Tomography, Optical Coherence ,Genome-Wide Association Study - Abstract
Purpose: This study aimed to identify the genetic defects in 2 families with autosomal recessive macular dystrophy with central cone involvement. Design: Case series. Participants: Two families and a cohort of 244 individuals with various inherited maculopathies and cone disorders. Methods: Genome-wide linkage analysis and exome sequencing were performed in 1 large family with 5 affected individuals. In addition, exome sequencing was performed in the proband of a second family. Subsequent analysis of the identified mutations in 244 patients was performed by Sanger sequencing or restriction enzyme digestion. The medical history of individuals carrying the MFSD8 variants was reviewed and additional ophthalmic examinations were performed, including electroretinography (ERG), multifocal ERG (mfERG), perimetry, optical coherence tomography (OCT), fundus autofluorescence, and fundus photography. Main Outcome Measures: MFSD8 variants, age at diagnosis, visual acuity, fundus appearance, color vision defects, visual field, ERG, mfERG, fundus autofluorescence, and OCT findings. Results: Compound heterozygous variants in MFSD8, a gene encoding a lysosomal transmembrane protein, were identified in 2 families with macular dystrophy with a normal or subnormal ERG, but reduced mfERG. In both families, a heterozygous missense variant p.Glu336Gln was identified, which was predicted to have a mild effect on the protein. In the first family, a protein-truncating variant (p.Glu381*) was identified on the other allele, and in the second family, a variant (c.1102G>C) was identified that results in a splicing defect leading to skipping of exon 11 (p.Lys333Lysfs*3). The p.Glu336Gln allele was found to be significantly enriched in patients with maculopathies and cone disorders (6/488) compared with ethnically matched controls (35/18 682; P < 0.0001), suggesting that it may act as a genetic modifier. Conclusions: In this study, we identified variants in MFSD8 as a novel cause of nonsyndromic autosomal recessive macular dystrophy with central cone involvement. Affected individuals showed no neurologic features typical for variant late-infantile neuronal ceroid lipofuscinosis (vLINCL), a severe and devastating multisystem lysosomal storage disease previously associated with mutations in MFSD8. We propose a genotype-phenotype model in which a combination of a severe and a mild variant cause nonsyndromic macular dystrophy with central cone involvement, and 2 severe mutations cause vLINCL. (C) 2015 by the American Academy of Ophthalmology.
- Published
- 2015
- Full Text
- View/download PDF
18. Autosomal dominant Ménétrier-like disease
- Author
-
Riccardo Sangermano, Erasmo Miele, Vito D. Corleto, Francesco Paolo D'Armiento, Maurizio Martini, Nicola Brunetti-Pierri, Pasquale Piccolo, Guido Rindi, Annamaria Staiano, Caterina Strisciuglio, Strisciuglio, Caterina, Corleto, Vd, Brunetti Pierri, N, Piccolo, P, Sangermano, R, Rindi, G, Martini, M, D'Armiento, Fp, Staiano, A, and Miele, E.
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Settore MED/12 - GASTROENTEROLOGIA ,Disease ,law.invention ,law ,medicine ,Humans ,Family ,Ménétrier disease ,Gastritis, Hypertrophic ,Pathological ,Gastrointestinal bleeding ,Polymerase chain reaction ,Genes, Dominant ,Hyperplasia ,business.industry ,Stomach ,Gastroenterology ,Case-control study ,Proteins ,Transforming Growth Factor alpha ,medicine.disease ,Transforming growth factor-α ,Pedigree ,Gastric Mucosa ,Metrier ,Case-Control Studies ,Child, Preschool ,Pediatrics, Perinatology and Child Health ,Immunohistochemistry ,Female ,Gastritis ,medicine.symptom ,business ,Transforming growth factor - Abstract
Background: Familial occurrence of Ménétrier disease is rare and has been reported only in few instances. Methods: Affected patients from a large pedigree were evaluated at the clinical, endoscopic, and pathological levels. Results: Affected members presented with gastropathy of variable severity but without protein loss. Endoscopy and pathology findings were consistent with Ménétrier disease; however, gastric transforming growth factor α (TGF-α) immunohistochemistry and real-time polymerase chain reaction showed no increase in TGF-α expression. Conclusions: We describe a unique, 4-generation pedigree with autosomal dominant gastropathy exhibiting the typical clinical, endoscopic, and pathological findings of Ménétrier-like disease, though in the absence of protein loss and with no increase in the levels of gastric TGF-α. Members of this family may be affected by a novel and previously unrecognised hereditary form of gastric hyperplasia. Copyright © 2012 by ESPGHAN and NASPGHAN.
- Published
- 2012
19. Coding and non-coding variants in the ciliopathy gene CFAP410 cause early-onset non-syndromic retinal degeneration.
- Author
-
Sangermano R, Gupta P, Price C, Han J, Navarro J, Condroyer C, Place EM, Antonio A, Mukai S, Zanlonghi X, Sahel JA, DiTroia S, O'Heir E, Duncan JL, Pierce EA, Zeitz C, Audo I, Huckfeldt RM, and Bujakowska KM
- Abstract
Inherited retinal degenerations are blinding genetic disorders characterized by high genetic and phenotypic heterogeneity. In this retrospective study, we describe sixteen families with early-onset non-syndromic retinal degenerations in which affected probands carried rare bi-allelic variants in CFAP410, a ciliary gene previously associated with recessive Jeune syndrome. We detected twelve variants, eight of which were novel, including c.373+91A>G, which led to aberrant splicing. To our knowledge this is the first likely pathogenic deep-intronic variant identified in this gene. Analysis of all reported and novel CFAP410 variants revealed no clear correlation between the severity of the CFAP410-associated phenotypes and the identified causal variants. This is supported by the fact that the frequently encountered missense variant p.(Arg73Pro), often found in syndromic cases, was also associated with non-syndromic retinal degeneration. This study expands the current knowledge of CFAP410-associated ciliopathy by enriching its mutational landscape and supports its association with non-syndromic retinal degeneration., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
20. Biallelic variation in the choline and ethanolamine transporter FLVCR1 underlies a severe developmental disorder spectrum.
- Author
-
Calame DG, Wong JH, Panda P, Nguyen DT, Leong NCP, Sangermano R, Patankar SG, Abdel-Hamid MS, AlAbdi L, Safwat S, Flannery KP, Dardas Z, Fatih JM, Murali C, Kannan V, Lotze TE, Herman I, Ammouri F, Rezich B, Efthymiou S, Alavi S, Murphy D, Firoozfar Z, Nasab ME, Bahreini A, Ghasemi M, Haridy NA, Goldouzi HR, Eghbal F, Karimiani EG, Begtrup A, Elloumi H, Srinivasan VM, Gowda VK, Du H, Jhangiani SN, Coban-Akdemir Z, Marafi D, Rodan L, Isikay S, Rosenfeld JA, Ramanathan S, Staton M, Oberg KC, Clark RD, Wenman C, Loughlin S, Saad R, Ashraf T, Male A, Tadros S, Boostani R, Abdel-Salam GMH, Zaki M, Mardi A, Hashemi-Gorji F, Abdalla E, Manzini MC, Pehlivan D, Posey JE, Gibbs RA, Houlden H, Alkuraya FS, Bujakowska K, Maroofian R, Lupski JR, and Nguyen LN
- Abstract
Purpose: FLVCR1 encodes a solute carrier protein implicated in heme, choline, and ethanolamine transport. Although Flvcr1
-/- mice exhibit skeletal malformations and defective erythropoiesis reminiscent of Diamond-Blackfan anemia (DBA), biallelic FLVCR1 variants in humans have previously only been linked to childhood or adult-onset ataxia, sensory neuropathy, and retinitis pigmentosa., Methods: We identified individuals with undiagnosed neurodevelopmental disorders and biallelic FLVCR1 variants through international data sharing and characterized the functional consequences of their FLVCR1 variants., Results: We ascertained 30 patients from 23 unrelated families with biallelic FLVCR1 variants and characterized a novel FLVCR1-related phenotype: severe developmental disorders with profound developmental delay, microcephaly (z-score -2.5 to -10.5), brain malformations, epilepsy, spasticity, and premature death. Brain malformations ranged from mild brain volume reduction to hydranencephaly. Severely affected patients share traits, including macrocytic anemia and skeletal malformations, with Flvcr1-/- mice and DBA. FLVCR1 variants significantly reduce choline and ethanolamine transport and/or disrupt mRNA splicing., Conclusion: These data demonstrate a broad FLVCR1-related phenotypic spectrum ranging from severe multiorgan developmental disorders resembling DBA to adult-onset neurodegeneration. Our study expands our understanding of Mendelian choline and ethanolamine disorders and illustrates the importance of anticipating a wide phenotypic spectrum for known disease genes and incorporating model organism data into genome analysis to maximize genetic testing yield., Competing Interests: Conflict of Interest James R. Lupski has stock ownership in 23andMe, is a paid consultant for Genome International, and is a coinventor on multiple US and European patents related to molecular diagnostics for inherited neuropathies, eye diseases, genomic disorders, and bacterial genomic fingerprinting. The Department of Molecular and Human Genetics at Baylor College of Medicine receives revenue from clinical genetic testing conducted at Baylor Genetics Laboratories. Amber Begtrup and Houda Elloumi are employees of GeneDx, LLC. All other authors declare no conflicts of interest., (Copyright © 2024 American College of Medical Genetics and Genomics. Published by Elsevier Inc. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
21. Targeted long-read sequencing enriches disease-relevant genomic regions of interest to provide complete Mendelian disease diagnostics.
- Author
-
Nakamichi K, Huey J, Sangermano R, Place EM, Bujakowska KM, Marra M, Everett LA, Yang P, Chao JR, Van Gelder RN, and Mustafi D
- Subjects
- Humans, Genetic Testing methods, Retinal Diseases genetics, Retinal Diseases diagnosis, Genome, Human, Genetic Diseases, Inborn genetics, Genetic Diseases, Inborn diagnosis, Genomics methods, Computational Biology methods, Sequence Analysis, DNA methods, Male, Haplotypes genetics, Female, High-Throughput Nucleotide Sequencing methods
- Abstract
Despite advances in sequencing technologies, a molecular diagnosis remains elusive in many patients with Mendelian disease. Current short-read clinical sequencing approaches cannot provide chromosomal phase information or epigenetic information without further sample processing, which is not routinely done and can result in an incomplete molecular diagnosis in patients. The ability to provide phased genetic and epigenetic information from a single sequencing run would improve the diagnostic rate of Mendelian conditions. Here, we describe targeted long-read sequencing of Mendelian disease genes (TaLon-SeqMD) using a real-time adaptive sequencing approach. Optimization of bioinformatic targeting enabled selective enrichment of multiple disease-causing regions of the human genome. Haplotype-resolved variant calling and simultaneous resolution of epigenetic base modification could be achieved in a single sequencing run. The TaLon-SeqMD approach was validated in a cohort of 18 individuals with previous genetic testing targeting 373 inherited retinal disease (IRD) genes, yielding the complete molecular diagnosis in each case. This approach was then applied in 2 IRD cases with inconclusive testing, which uncovered noncoding and structural variants that were difficult to characterize by standard short-read sequencing. Overall, these results demonstrate TaLon-SeqMD as an approach to provide rapid phased-variant calling to provide the molecular basis of Mendelian diseases.
- Published
- 2024
- Full Text
- View/download PDF
22. Substitution of a single non-coding nucleotide upstream of TMEM216 causes non-syndromic retinitis pigmentosa and is associated with reduced TMEM216 expression.
- Author
-
Malka S, Biswas P, Berry AM, Sangermano R, Ullah M, Lin S, D'Antonio M, Jestin A, Jiao X, Quinodoz M, Sullivan L, Gardner JC, Place EM, Michaelides M, Kaminska K, Mahroo OA, Schiff E, Wright G, Cancellieri F, Vaclavik V, Santos C, Rehman AU, Mehrotra S, Azhar Baig HM, Iqbal M, Ansar M, Santos LC, Sousa AB, Tran VH, Matsui H, Bhatia A, Naeem MA, Akram SJ, Akram J, Riazuddin S, Ayuso C, Pierce EA, Hardcastle AJ, Riazuddin SA, Frazer KA, Hejtmancik JF, Rivolta C, Bujakowska KM, Arno G, Webster AR, and Ayyagari R
- Subjects
- Adult, Child, Child, Preschool, Female, Humans, Male, Young Adult, Alleles, Haplotypes, Heterozygote, Homozygote, Membrane Proteins genetics, Phenotype, Pedigree, Polymorphism, Single Nucleotide, Retinitis Pigmentosa genetics, Retinitis Pigmentosa pathology
- Abstract
Genome analysis of individuals affected by retinitis pigmentosa (RP) identified two rare nucleotide substitutions at the same genomic location on chromosome 11 (g.61392563 [GRCh38]), 69 base pairs upstream of the start codon of the ciliopathy gene TMEM216 (c.-69G>A, c.-69G>T [GenBank: NM_001173991.3]), in individuals of South Asian and African ancestry, respectively. Genotypes included 71 homozygotes and 3 mixed heterozygotes in trans with a predicted loss-of-function allele. Haplotype analysis showed single-nucleotide variants (SNVs) common across families, suggesting ancestral alleles within the two distinct ethnic populations. Clinical phenotype analysis of 62 available individuals from 49 families indicated a similar clinical presentation with night blindness in the first decade and progressive peripheral field loss thereafter. No evident systemic ciliopathy features were noted. Functional characterization of these variants by luciferase reporter gene assay showed reduced promotor activity. Nanopore sequencing confirmed the lower transcription of the TMEM216 c.-69G>T allele in blood-derived RNA from a heterozygous carrier, and reduced expression was further recapitulated by qPCR, using both leukocytes-derived RNA of c.-69G>T homozygotes and total RNA from genome-edited hTERT-RPE1 cells carrying homozygous TMEM216 c.-69G>A. In conclusion, these variants explain a significant proportion of unsolved cases, specifically in individuals of African ancestry, suggesting that reduced TMEM216 expression might lead to abnormal ciliogenesis and photoreceptor degeneration., Competing Interests: Declaration of interests All the authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
23. Clinician-Driven Reanalysis of Exome Sequencing Data From Patients With Inherited Retinal Diseases.
- Author
-
Surl D, Won D, Lee ST, Lee CS, Lee J, Lim HT, Chung SA, Song WK, Kim M, Kim SS, Shin S, Choi JR, Sangermano R, Byeon SH, Bujakowska KM, and Han J
- Subjects
- Humans, Male, Female, Adult, Prospective Studies, Middle Aged, Republic of Korea, Genetic Testing methods, Genetic Testing statistics & numerical data, Adolescent, Young Adult, Child, High-Throughput Nucleotide Sequencing methods, Computational Biology methods, Exome Sequencing methods, Retinal Diseases genetics, Retinal Diseases diagnosis
- Abstract
Importance: Despite advances in next-generation sequencing (NGS), a significant proportion of patients with inherited retinal disease (IRD) remain undiagnosed after initial genetic testing. Exome sequencing (ES) reanalysis in the clinical setting has been suggested as one method for improving diagnosis of IRD., Objective: To investigate the association of clinician-led reanalysis of ES data, which incorporates updated clinical information and comprehensive bioinformatic analysis, with the diagnostic yield in a cohort of patients with IRDs in Korea., Design, Setting, and Participants: This was a multicenter prospective cohort study involving 264 unrelated patients with IRDs, conducted in Korea between March 2018 and February 2020. Comprehensive ophthalmologic examinations and ES analyses were performed, and ES data were reanalyzed by an IRD specialist for single nucleotide variants, copy number variants, mobile element insertions, and mitochondrial variants. Data were analyzed from March to July 2023., Main Outcomes and Measures: Diagnostic rate of conventional bioinformatic analysis and clinician-driven ES reanalysis., Results: A total of 264 participants (151 [57.2%] male; mean [SD] age at genetic testing, 33.6 [18.9] years) were enrolled, including 129 patients (48.9%) with retinitis pigmentosa and 26 patients (9.8%) with Stargardt disease or macular dystrophy. Initial bioinformatic analysis diagnosed 166 patients (62.9%). Clinician-driven reanalysis identified the molecular cause of diseases in an additional 22 patients, corresponding to an 8.3-percentage point increase in diagnostic rate. Key factors associated with new molecular diagnoses included clinical phenotype updates (4 patients) and detection of previously overlooked variation, such as structural variants (9 patients), mitochondrial variants (3 patients), filtered or not captured variants (4 patients), and noncanonical splicing variants (2 patients). Among the 22 patients, variants in 7 patients (31.8%) were observed in the initial analysis but not reported to patients, while those in the remaining 15 patients (68.2%) were newly detected by the ES reanalysis., Conclusions and Relevance: In this cohort study, clinician-centered reanalysis of ES data was associated with improved molecular diagnostic yields in patients with IRD. This approach is important for uncovering missed genetic causes of retinal disease.
- Published
- 2024
- Full Text
- View/download PDF
24. Biallelic variation in the choline and ethanolamine transporter FLVCR1 underlies a pleiotropic disease spectrum from adult neurodegeneration to severe developmental disorders.
- Author
-
Calame DG, Wong JH, Panda P, Nguyen DT, Leong NCP, Sangermano R, Patankar SG, Abdel-Hamid M, AlAbdi L, Safwat S, Flannery KP, Dardas Z, Fatih JM, Murali C, Kannan V, Lotze TE, Herman I, Ammouri F, Rezich B, Efthymiou S, Alavi S, Murphy D, Firoozfar Z, Nasab ME, Bahreini A, Ghasemi M, Haridy NA, Goldouzi HR, Eghbal F, Karimiani EG, Srinivasan VM, Gowda VK, Du H, Jhangiani SN, Coban-Akdemir Z, Marafi D, Rodan L, Isikay S, Rosenfeld JA, Ramanathan S, Staton M, Kerby C Oberg, Clark RD, Wenman C, Loughlin S, Saad R, Ashraf T, Male A, Tadros S, Boostani R, Abdel-Salam GMH, Zaki M, Abdalla E, Manzini MC, Pehlivan D, Posey JE, Gibbs RA, Houlden H, Alkuraya FS, Bujakowska K, Maroofian R, Lupski JR, and Nguyen LN
- Abstract
FLVCR1 encodes Feline leukemia virus subgroup C receptor 1 (FLVCR1), a solute carrier (SLC) transporter within the Major Facilitator Superfamily. FLVCR1 is a widely expressed transmembrane protein with plasma membrane and mitochondrial isoforms implicated in heme, choline, and ethanolamine transport. While Flvcr1 knockout mice die in utero with skeletal malformations and defective erythropoiesis reminiscent of Diamond-Blackfan anemia, rare biallelic pathogenic FLVCR1 variants are linked to childhood or adult-onset neurodegeneration of the retina, spinal cord, and peripheral nervous system. We ascertained from research and clinical exome sequencing 27 individuals from 20 unrelated families with biallelic ultra-rare missense and predicted loss-of-function (pLoF) FLVCR1 variant alleles. We characterize an expansive FLVCR1 phenotypic spectrum ranging from adult-onset retinitis pigmentosa to severe developmental disorders with microcephaly, reduced brain volume, epilepsy, spasticity, and premature death. The most severely affected individuals, including three individuals with homozygous pLoF variants, share traits with Flvcr1 knockout mice and Diamond-Blackfan anemia including macrocytic anemia and congenital skeletal malformations. Pathogenic FLVCR1 missense variants primarily lie within transmembrane domains and reduce choline and ethanolamine transport activity compared with wild-type FLVCR1 with minimal impact on FLVCR1 stability or subcellular localization. Several variants disrupt splicing in a mini-gene assay which may contribute to genotype-phenotype correlations. Taken together, these data support an allele-specific gene dosage model in which phenotypic severity reflects residual FLVCR1 activity. This study expands our understanding of Mendelian disorders of choline and ethanolamine transport and demonstrates the importance of choline and ethanolamine in neurodevelopment and neuronal homeostasis., Competing Interests: Potential Conflict of Interest J.R.L. has stock ownership in 23andMe, is a paid consultant for Genome International, and is a co-inventor on multiple United States and European patents related to molecular diagnostics for inherited neuropathies, eye diseases, genomic disorders, and bacterial genomic fingerprinting. The Department of Molecular and Human Genetics at Baylor College of Medicine receives revenue from clinical genetic testing conducted at Baylor Genetics (BG) Laboratories. Other authors have no potential conflicts to disclose.
- Published
- 2024
- Full Text
- View/download PDF
25. Non-syndromic Retinal Degeneration Caused by Pathogenic Variants in Joubert Syndrome Genes.
- Author
-
Sangermano R, Galdikaité-Braziené E, and Bujakowska KM
- Subjects
- Humans, Retina pathology, Cerebellum pathology, Mutation, Pedigree, Retinal Degeneration genetics, Retinal Degeneration pathology, Abnormalities, Multiple genetics, Abnormalities, Multiple pathology, Kidney Diseases, Cystic genetics, Eye Abnormalities genetics, Eye Abnormalities pathology
- Abstract
Inherited retinal degenerations (IRDs) are a group of genetic disorders characterized by progressive dysfunction and loss of photoreceptors. IRDs are classified as non-syndromic or syndromic, depending on whether retinal degeneration manifests alone or in combination with other associated symptoms. Joubert syndrome (JBTS) is a genetically and clinically heterogeneous disorder affecting the central nervous system and other organs and tissues, including the neuroretina. To date, 39 genes have been associated with JBTS, a majority of which encode structural or functional components of the primary cilium, a specialized sensory organelle present in most post-mitotic cells, including photoreceptors. The use of whole exome and IRD panel next-generation sequencing in routine diagnostics of non-syndromic IRD cases led to the discovery of pathogenic variants in JBTS genes that cause photoreceptor loss without other syndromic features. Here, we recapitulate these findings, describing the JBTS gene defects leading to non-syndromic IRDs., (© 2023. The Author(s), under exclusive license to Springer Nature Switzerland AG.)
- Published
- 2023
- Full Text
- View/download PDF
26. Identification of a novel large multigene deletion and a frameshift indel in PDE6B as the underlying cause of early-onset recessive rod-cone degeneration.
- Author
-
Sangermano R, Biswas P, Sullivan LS, Place EM, Borooah S, Straubhaar J, Pierce EA, Daiger SP, Bujakowska KM, and Ayaggari R
- Subjects
- Humans, Frameshift Mutation genetics, Blindness genetics, INDEL Mutation, Pedigree, Mutation, Cyclic Nucleotide Phosphodiesterases, Type 6 genetics, Retinal Degeneration genetics, Night Blindness genetics
- Abstract
A family, with two affected identical twins with early-onset recessive inherited retinal degeneration, was analyzed to determine the underlying genetic cause of pathology. Exome sequencing revealed a rare and previously reported causative variant (c.1923_1969delinsTCTGGG; p.Asn643Glyfs*29) in the PDE6B gene in the affected twins and their unaffected father. Further investigation, using genome sequencing, identified a novel ∼7.5-kb deletion (Chr 4:670,405-677,862del) encompassing the ATP5ME gene, part of the 5' UTR of MYL5 , and a 378-bp (Chr 4:670,405-670,782) region from the 3' UTR of PDE6B in the affected twins and their unaffected mother. Both variants segregated with disease in the family. Analysis of the relative expression of PDE6B , in peripheral blood cells, also revealed a significantly lower level of PDE6B transcript in affected siblings compared to a normal control. PDE6B is associated with recessive rod-cone degeneration and autosomal dominant congenital stationary night blindness. Ophthalmic evaluation of these patients showed night blindness, fundus abnormalities, and peripheral vision loss, which are consistent with PDE6B -associated recessive retinal degeneration. These findings suggest that the loss of PDE6B transcript resulting from the compound heterozygous pathogenic variants is the underlying cause of recessive rod-cone degeneration in the study family., (© 2022 Sangermano et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2022
- Full Text
- View/download PDF
27. Novel RCBTB1 variants causing later-onset non-syndromic retinal dystrophy with macular chorioretinal atrophy.
- Author
-
Catomeris AJ, Ballios BG, Sangermano R, Wagner NE, Comander JI, Pierce EA, Place EM, Bujakowska KM, and Huckfeldt RM
- Subjects
- Atrophy, Canada ethnology, Guanine Nucleotide Exchange Factors genetics, Humans, Pedigree, Phenotype, Retrospective Studies, Macular Degeneration genetics, Retinal Dystrophies diagnosis, Retinal Dystrophies genetics
- Abstract
Background: Variants in RCBTB1 were recently described to cause a retinal dystrophy with only eight families described to date and a predominant phenotype of macular atrophy and peripheral reticular degeneration. Here, we further evaluate the genotypic and phenotypic characteristics of biallelic RCBTB1 -associated retinal dystrophy in a North American clinic population., Methods: A retrospective analysis of genetic and clinical features was performed in individuals with biallelic variants in RCBTB1 ., Results: Three unrelated individuals of French-Canadian descent with rare biallelic RCBTB1 variants were identified. All individuals shared a novel p.(Ser342Leu) missense variant; one patient was homozygous whereas the other two each possessed a second unique novel variant p.(Gln120*) and p.(Pro224Leu). All three had macula-predominant disease with symptom onset in the fifth decade of life., Conclusion: This report adds to the genetic diversity of RCBTB1 -associated disease. These cases confirm the later-onset, relative to many other retinal dystrophies, and macular focus of disease described in most cases to-date. They are thus a reminder of considering hereditary disease in the differential for later-onset macular atrophy.
- Published
- 2022
- Full Text
- View/download PDF
28. Broadening INPP5E phenotypic spectrum: detection of rare variants in syndromic and non-syndromic IRD.
- Author
-
Sangermano R, Deitch I, Peter VG, Ba-Abbad R, Place EM, Zampaglione E, Wagner NE, Fulton AB, Coutinho-Santos L, Rosin B, Dunet V, AlTalbishi A, Banin E, Sousa AB, Neves M, Larson A, Quinodoz M, Michaelides M, Ben-Yosef T, Pierce EA, Rivolta C, Webster AR, Arno G, Sharon D, Huckfeldt RM, and Bujakowska KM
- Abstract
Pathogenic variants in INPP5E cause Joubert syndrome (JBTS), a ciliopathy with retinal involvement. However, despite sporadic cases in large cohort sequencing studies, a clear association with non-syndromic inherited retinal degenerations (IRDs) has not been made. We validate this association by reporting 16 non-syndromic IRD patients from ten families with bi-allelic mutations in INPP5E. Additional two patients showed early onset IRD with limited JBTS features. Detailed phenotypic description for all probands is presented. We report 14 rare INPP5E variants, 12 of which have not been reported in previous studies. We present tertiary protein modeling and analyze all INPP5E variants for deleteriousness and phenotypic correlation. We observe that the combined impact of INPP5E variants in JBTS and non-syndromic IRD patients does not reveal a clear genotype-phenotype correlation, suggesting the involvement of genetic modifiers. Our study cements the wide phenotypic spectrum of INPP5E disease, adding proof that sequence defects in this gene can lead to early-onset non-syndromic IRD.
- Published
- 2021
- Full Text
- View/download PDF
29. Lactoferrin and oral pathologies: a therapeutic treatment.
- Author
-
Rosa L, Lepanto MS, Cutone A, Ianiro G, Pernarella S, Sangermano R, Musci G, Ottolenghi L, and Valenti P
- Subjects
- Administration, Topical, Animals, Anti-Bacterial Agents administration & dosage, Anti-Bacterial Agents metabolism, Bacteria drug effects, Biofilms drug effects, Humans, Inflammation drug therapy, Inflammation metabolism, Inflammation pathology, Lactoferrin administration & dosage, Mouth microbiology, Mouth pathology, Anti-Bacterial Agents pharmacology, Lactoferrin metabolism, Mouth drug effects
- Abstract
The oral cavity is a non-uniform, extraordinary environment characterized by mucosal, epithelial, abiotic surfaces and secretions as saliva. Aerobic and anaerobic commensal and pathogenic microorganisms colonize the tongue, teeth, jowl, gingiva, and periodontium. Commensals exert an important role in host defenses, while pathogenic microorganisms can nullify this protective function causing oral and systemic diseases. Every day, 750-1000 mL of saliva, containing several host defense constituents including lactoferrin (Lf), are secreted and swallowed. Lf is a multifunctional iron-chelating cationic glycoprotein of innate immunity. Depending on, or regardless of its iron-binding ability, Lf exerts bacteriostatic, bactericidal, antibiofilm, antioxidant, antiadhesive, anti-invasive, and anti-inflammatory activities. Here, we report the protective role of Lf in different oral pathologies, such as xerostomia, halitosis, alveolar or maxillary bone damage, gingivitis, periodontitis, and black stain. Unlike antibiotic therapy, which is ineffective against bacteria that are within a biofilm, adherent, or intracellular, the topical administration of Lf, through its simultaneous activity against microbial replication, biofilms, adhesion, and invasiveness, as well as inflammation, has been proven to be efficient in the treatment of all known oral pathologies without any adverse effects.
- Published
- 2021
- Full Text
- View/download PDF
30. A combined RNA-seq and whole genome sequencing approach for identification of non-coding pathogenic variants in single families.
- Author
-
Bronstein R, Capowski EE, Mehrotra S, Jansen AD, Navarro-Gomez D, Maher M, Place E, Sangermano R, Bujakowska KM, Gamm DM, and Pierce EA
- Subjects
- Child, Female, Gene Expression Profiling, Humans, Male, Pedigree, Exome Sequencing, Genetic Markers, Genetic Variation, Genome, Human, RNA-Seq methods, Retinal Degeneration genetics, Retinal Degeneration pathology, Whole Genome Sequencing methods
- Abstract
Inherited retinal degenerations (IRDs) are at the focus of current genetic therapeutic advancements. For a genetic treatment such as gene therapy to be successful, an accurate genetic diagnostic is required. Genetic diagnostics relies on the assessment of the probability that a given DNA variant is pathogenic. Non-coding variants present a unique challenge for such assessments as compared to coding variants. For one, non-coding variants are present at much higher number in the genome than coding variants. In addition, our understanding of the rules that govern the non-coding regions of the genome is less complete than our understanding of the coding regions. Methods that allow for both the identification of candidate non-coding pathogenic variants and their functional validation may help overcome these caveats allowing for a greater number of patients to benefit from advancements in genetic therapeutics. We present here an unbiased approach combining whole genome sequencing (WGS) with patient-induced pluripotent stem cell (iPSC)-derived retinal organoids (ROs) transcriptome analysis. With this approach, we identified and functionally validated a novel pathogenic non-coding variant in a small family with a previously unresolved genetic diagnosis., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
- Full Text
- View/download PDF
31. In or Out? New Insights on Exon Recognition through Splice-Site Interdependency.
- Author
-
Khan M, Cornelis SS, Sangermano R, Post IJM, Groesbeek AJ, Amsu J, Gilissen C, Garanto A, Collin RWJ, and Cremers FPM
- Subjects
- ATP-Binding Cassette Transporters genetics, ATP-Binding Cassette Transporters metabolism, Dystrophin genetics, Dystrophin metabolism, HEK293 Cells, Humans, Membrane Proteins genetics, Membrane Proteins metabolism, RNA Splicing, Exons, RNA Splice Sites
- Abstract
Noncanonical splice-site mutations are an important cause of inherited diseases. Based on in vitro and stem-cell-based studies, some splice-site variants show a stronger splice defect than expected based on their predicted effects, suggesting that other sequence motifs influence the outcome. We investigated whether splice defects due to human-inherited-disease-associated variants in noncanonical splice-site sequences in ABCA4 , DMD , and TMC1 could be rescued by strengthening the splice site on the other side of the exon. Noncanonical 5'- and 3'-splice-site variants were selected. Rescue variants were introduced based on an increase in predicted splice-site strength, and the effects of these variants were analyzed using in vitro splice assays in HEK293T cells. Exon skipping due to five variants in noncanonical splice sites of exons in ABCA4 , DMD , and TMC1 could be partially or completely rescued by increasing the predicted strengths of the other splice site of the same exon. We named this mechanism "splicing interdependency", and it is likely based on exon recognition by splicing machinery. Awareness of this interdependency is of importance in the classification of noncanonical splice-site variants associated with disease and may open new opportunities for treatments.
- Published
- 2020
- Full Text
- View/download PDF
32. Late-Onset Stargardt Disease Due to Mild, Deep-Intronic ABCA4 Alleles.
- Author
-
Runhart EH, Valkenburg D, Cornelis SS, Khan M, Sangermano R, Albert S, Bax NM, Astuti GDN, Gilissen C, Pott JR, Verheij JBGM, Blokland EAW, Cremers FPM, van den Born LI, and Hoyng CB
- Subjects
- Aged, Alleles, Female, Gene Frequency, Genetic Variation, Humans, Kaplan-Meier Estimate, Male, Middle Aged, Visual Acuity, ATP-Binding Cassette Transporters genetics, Stargardt Disease genetics
- Abstract
Purpose: To investigate the role of two deep-intronic ABCA4 variants, that showed a mild splice defect in vitro and can occur on the same allele as the low penetrant c.5603A>T, in Stargardt disease (STGD1)., Methods: Ophthalmic data were assessed of 18 STGD1 patients who harbored c.769-784C>T or c.4253+43G>A in combination with a severe ABCA4 variant. Subjects carrying c.[769-784C>T; 5603A>T] were clinically compared with a STGD1 cohort previously published carrying c.5603A>T noncomplex. We calculated the penetrances of the intronic variants using ABCA4 allele frequency data of the general population and investigated the effect of c.769-784C>T on splicing in photoreceptor progenitor cells (PPCs)., Results: Mostly, late-onset, foveal-sparing STGD1 was observed among subjects harboring c.769-784C>T or c.4253+43G>A (median age of onset, 54.5 and 52.0 years, respectively). However, ages of onset, phenotypes in fundo, and visual acuity courses varied widely. No significant clinical differences were observed between the c.[769-784C>T; 5603A>T] cohort and the c.4253+43G>A or the c.5603A>T cohort. The penetrances of c.769-784C>T (20.5%-39.6%) and c.4253+43G>A (35.8%-43.1%) were reduced, when not considering the effect of yet unidentified or known factors in cis, such as c.5603A>T (identified in 7/7 probands with c.769-784C>T; 1/8 probands with c.4253+43G>A). Variant c.769-784C>T resulted in a pseudo-exon insertion in 15% of the total mRNA (i.e., ∼30% of the c.769-784C>T allele alone)., Conclusions: Two mild intronic ABCA4 variants could further explain missing heritability in late-onset STGD1, distinguishing it from AMD. The observed clinical variability and calculated reduced penetrance urge research into modifiers within and outside of the ABCA4 gene.
- Published
- 2019
- Full Text
- View/download PDF
33. The treatment of black stain associated with of iron metabolism disorders with lactoferrin: a litterature search and two case studies.
- Author
-
Sangermano R, Pernarella S, Straker M, Lepanto MS, Rosa L, Cutone A, Valenti P, and Ottolenghi L
- Subjects
- Adult, Child, Dental Caries diagnosis, Dental Plaque diagnosis, Female, Humans, Iron Metabolism Disorders diagnosis, Male, Pregnancy, Saliva metabolism, Tooth Discoloration drug therapy, Anti-Infective Agents administration & dosage, Dental Caries diet therapy, Iron Metabolism Disorders drug therapy, Lactoferrin administration & dosage, Tooth Discoloration diagnosis
- Abstract
Among the various pathologies of the oral cavity, the formation of "unsightly black spots" on the surface of the tooth, universally known as Black Stain (BS) has recently been acquiring more interest. Usually BS is typically found in individuals in prepubertal age, even though it has been identified in adults associated with microbial exchange and / or with iron metabolism disorders. Microbial exchange concerns the possible exchange of bacteria between family members which can take place directly, through effusions, or indirectly, through brushes, cutlery or glasses. For this reason, it is recommended that toothbrushes of family members not be left damp and in contact with each other. The bathroom, being a warm-humid environment, is in fact an optimal habitat for microbial proliferation. Of specific importance in BS is the accumulation of iron in tissues and secretions which, together with chromogenic bacteria, are the primary cause of this pathology. In fact, among the metabolic products synthesized by bacteria in the oral cavity, hydrogen sulfide is of considerable interest, since upon reacting with iron available in saliva, in pathological conditions (iron metabolism disorders), it forms black precipitates consisting of ferric sulfide. These precipitates bind to the surface of the teeth, tending to form a stria that usually follows the contour of the gingiva, with an unsightly and variable chromatic intensity. In physiological situations, iron homeostasis is defined as the state of equilibrium between iron present in tissues and in secretions and that which is present in the circulation. Instead, in pathological conditions, defined as iron metabolism disorders, there is an accumulation of iron in tissues and secretions and a lack of it in the circulation. It is also important to remember that subjects affected by BS are more protected from carious processes than healthy subjects, probably due to a significant predominance of chromogenic bacteria compared to those responsible for caries. It should also be remembered that in young subjects BS tends to regress with pubertal development and the transition to adult life. In any case, using common professional hygiene procedures, it is possible to remove BS as well as plaque and tartar deposits. In particular, with ultrasonic scalers, polishing pastes and powders carried by air and water jets, the surfaces of the teeth can be restored to their natural healthy state. All the techniques for removing the precipitates, are not enough however, to fix and permanently eradicate their appearance, as these precipitates last only for short periods and recur very frequently. Due to the frequent recurrences, new oral microbiota control therapies are emerging; among these the use of lactoferrin (Lf) in the dental field and particularly in the treatment of BS appears to be very promising. Taken togheter, here the effect of Lf in subjects affected by BS has been investigated.
- Published
- 2019
- Full Text
- View/download PDF
34. ABCA4-associated disease as a model for missing heritability in autosomal recessive disorders: novel noncoding splice, cis-regulatory, structural, and recurrent hypomorphic variants.
- Author
-
Bauwens M, Garanto A, Sangermano R, Naessens S, Weisschuh N, De Zaeytijd J, Khan M, Sadler F, Balikova I, Van Cauwenbergh C, Rosseel T, Bauwens J, De Leeneer K, De Jaegere S, Van Laethem T, De Vries M, Carss K, Arno G, Fakin A, Webster AR, de Ravel de l'Argentière TJL, Sznajer Y, Vuylsteke M, Kohl S, Wissinger B, Cherry T, Collin RWJ, Cremers FPM, Leroy BP, and De Baere E
- Subjects
- Adult, Alleles, Cohort Studies, Exons genetics, Female, Gene Frequency, HEK293 Cells, Humans, Introns genetics, Male, Middle Aged, Mutation genetics, Oligonucleotides, Antisense pharmacology, Pedigree, Phenotype, Retinal Dystrophies pathology, ATP-Binding Cassette Transporters genetics, Genes, Recessive genetics, Oligonucleotides, Antisense genetics, Retinal Dystrophies genetics
- Abstract
Purpose: ABCA4-associated disease, a recessive retinal dystrophy, is hallmarked by a large proportion of patients with only one pathogenic ABCA4 variant, suggestive for missing heritability., Methods: By locus-specific analysis of ABCA4, combined with extensive functional studies, we aimed to unravel the missing alleles in a cohort of 67 patients (p), with one (p = 64) or no (p = 3) identified coding pathogenic variants of ABCA4., Results: We identified eight pathogenic (deep-)intronic ABCA4 splice variants, of which five are novel and six structural variants, four of which are novel, including two duplications. Together, these variants account for the missing alleles in 40.3% of patients. Furthermore, two novel variants with a putative cis-regulatory effect were identified. The common hypomorphic variant c.5603A>T p.(Asn1868Ile) was found as a candidate second allele in 43.3% of patients. Overall, we have elucidated the missing heritability in 83.6% of our cohort. In addition, we successfully rescued three deep-intronic variants using antisense oligonucleotide (AON)-mediated treatment in HEK 293-T cells and in patient-derived fibroblast cells., Conclusion: Noncoding pathogenic variants, novel structural variants, and a common hypomorphic allele of the ABCA4 gene explain the majority of unsolved cases with ABCA4-associated disease, rendering this retinopathy a model for missing heritability in autosomal recessive disorders.
- Published
- 2019
- Full Text
- View/download PDF
35. Deep-intronic ABCA4 variants explain missing heritability in Stargardt disease and allow correction of splice defects by antisense oligonucleotides.
- Author
-
Sangermano R, Garanto A, Khan M, Runhart EH, Bauwens M, Bax NM, van den Born LI, Khan MI, Cornelis SS, Verheij JBGM, Pott JR, Thiadens AAHJ, Klaver CCW, Puech B, Meunier I, Naessens S, Arno G, Fakin A, Carss KJ, Raymond FL, Webster AR, Dhaenens CM, Stöhr H, Grassmann F, Weber BHF, Hoyng CB, De Baere E, Albert S, Collin RWJ, and Cremers FPM
- Subjects
- Adolescent, Adult, Aged, Child, Exons genetics, HEK293 Cells, Humans, Introns genetics, Middle Aged, Mutation genetics, Oligonucleotides, Antisense pharmacology, Pedigree, Polymorphism, Single Nucleotide genetics, RNA Splicing genetics, Stargardt Disease pathology, Young Adult, ATP-Binding Cassette Transporters genetics, Oligonucleotides, Antisense genetics, Protein Isoforms genetics, Stargardt Disease genetics
- Abstract
Purpose: Using exome sequencing, the underlying variants in many persons with autosomal recessive diseases remain undetected. We explored autosomal recessive Stargardt disease (STGD1) as a model to identify the missing heritability., Methods: Sequencing of ABCA4 was performed in 8 STGD1 cases with one variant and p.Asn1868Ile in trans, 25 cases with one variant, and 3 cases with no ABCA4 variant. The effect of intronic variants was analyzed using in vitro splice assays in HEK293T cells and patient-derived fibroblasts. Antisense oligonucleotides were used to correct splice defects., Results: In 24 of the probands (67%), one known and five novel deep-intronic variants were found. The five novel variants resulted in messenger RNA pseudoexon inclusions, due to strengthening of cryptic splice sites or by disrupting a splicing silencer motif. Variant c.769-784C>T showed partial insertion of a pseudoexon and was found in cis with c.5603A>T (p.Asn1868Ile), so its causal role could not be fully established. Variant c.4253+43G>A resulted in partial skipping of exon 28. Remarkably, antisense oligonucleotides targeting the aberrant splice processes resulted in (partial) correction of all splicing defects., Conclusion: Our data demonstrate the importance of assessing noncoding variants in genetic diseases, and show the great potential of splice modulation therapy for deep-intronic variants.
- Published
- 2019
- Full Text
- View/download PDF
36. The Common ABCA4 Variant p.Asn1868Ile Shows Nonpenetrance and Variable Expression of Stargardt Disease When Present in trans With Severe Variants.
- Author
-
Runhart EH, Sangermano R, Cornelis SS, Verheij JBGM, Plomp AS, Boon CJF, Lugtenberg D, Roosing S, Bax NM, Blokland EAW, Jacobs-Camps MHM, van der Velde-Visser SD, Pott JR, Rohrschneider K, Thiadens AAHJ, Klaver CCW, van den Born LI, Hoyng CB, and Cremers FPM
- Subjects
- Adult, Age of Onset, Aged, Electroretinography, Female, Fluorescein Angiography, Gene Frequency, Genetic Complementation Test, Humans, Macular Degeneration diagnosis, Macular Degeneration genetics, Macular Degeneration physiopathology, Male, Middle Aged, Pedigree, Penetrance, Retrospective Studies, Sequence Analysis, DNA, Siblings, Stargardt Disease, Tomography, Optical Coherence, Visual Acuity physiology, ATP-Binding Cassette Transporters genetics, Macular Degeneration congenital, Mutation, Polymorphism, Single Nucleotide
- Abstract
Purpose: To assess the occurrence and the disease expression of the common p.Asn1868Ile variant in patients with Stargardt disease (STGD1) harboring known, monoallelic causal ABCA4 variants., Methods: The coding and noncoding regions of ABCA4 were sequenced in 67 and 63 STGD1 probands respectively, harboring monoallelic ABCA4 variants. In case p.Asn1868Ile was detected, segregation analysis was performed whenever possible. Probands and affected siblings harboring p.Asn1868Ile without additional variants in cis were clinically evaluated retrospectively. Two asymptomatic siblings carrying the same ABCA4 variants as their probands were clinically examined. The penetrance of p.Asn1868Ile was calculated using allele frequency data of ABCA4 variants in non-Finnish European individuals., Results: The p.Asn1868Ile variant was found in cis with known variants in 14/67 probands. In 27/67 probands, we identified p.Asn1868Ile without additional variants in cis, in combination with known, mainly severe ABCA4 variants. In 23/27 probands, the trans configuration was established. Among 27 probands and 6/7 STGD1 siblings carrying p.Asn1868Ile, 42% manifested late-onset disease (>44 years). We additionally identified four asymptomatic relatives carrying a combination of a severe variant and p.Asn1868Ile; ophthalmologic examination in two persons did not reveal STGD1. Based on ABCA4 allele frequency data, we conservatively estimated the penetrance of p.Asn1868Ile, when present in trans with a severe variant, to be below 5%., Conclusions: A significant fraction of genetically unexplained STGD1 cases carries p.Asn1868Ile as a second variant. Our findings suggest exceptional differences in disease expression or even nonpenetrance of this ABCA4 variant, pointing toward an important role for genetic or environmental modifiers in STGD1.
- Published
- 2018
- Full Text
- View/download PDF
37. Identification and Rescue of Splice Defects Caused by Two Neighboring Deep-Intronic ABCA4 Mutations Underlying Stargardt Disease.
- Author
-
Albert S, Garanto A, Sangermano R, Khan M, Bax NM, Hoyng CB, Zernant J, Lee W, Allikmets R, Collin RWJ, and Cremers FPM
- Subjects
- Alleles, Base Sequence, Computer Simulation, Exons genetics, Humans, Macular Degeneration genetics, Oligonucleotides, Antisense pharmacology, Photoreceptor Cells, Vertebrate drug effects, Photoreceptor Cells, Vertebrate metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Stargardt Disease, ATP-Binding Cassette Transporters genetics, Introns genetics, Macular Degeneration congenital, Mutation genetics, RNA Splice Sites genetics
- Abstract
Sequence analysis of the coding regions and splice site sequences in inherited retinal diseases is not able to uncover ∼40% of the causal variants. Whole-genome sequencing can identify most of the non-coding variants, but their interpretation is still very challenging, in particular when the relevant gene is expressed in a tissue-specific manner. Deep-intronic variants in ABCA4 have been associated with autosomal-recessive Stargardt disease (STGD1), but the exact pathogenic mechanism is unknown. By generating photoreceptor precursor cells (PPCs) from fibroblasts obtained from individuals with STGD1, we demonstrated that two neighboring deep-intronic ABCA4 variants (c.4539+2001G>A and c.4539+2028C>T) result in a retina-specific 345-nt pseudoexon insertion (predicted protein change: p.Arg1514Leufs
∗ 36), likely due to the creation of exonic enhancers. Administration of antisense oligonucleotides (AONs) targeting the 345-nt pseudoexon can significantly rescue the splicing defect observed in PPCs of two individuals with these mutations. Intriguingly, an AON that is complementary to c.4539+2001G>A rescued the splicing defect only in PPCs derived from an individual with STGD1 with this but not the other mutation, demonstrating the high specificity of AONs. In addition, a single AON molecule rescued splicing defects associated with different neighboring mutations, thereby providing new strategies for the treatment of persons with STGD1. As many genes associated with human genetic conditions are expressed in specific tissues and pre-mRNA splicing may also rely on organ-specific factors, our approach to investigate and treat splicing variants using differentiated cells derived from individuals with STGD1 can be applied to any tissue of interest., (Copyright © 2018 American Society of Human Genetics. All rights reserved.)- Published
- 2018
- Full Text
- View/download PDF
38. ABCA4 midigenes reveal the full splice spectrum of all reported noncanonical splice site variants in Stargardt disease.
- Author
-
Sangermano R, Khan M, Cornelis SS, Richelle V, Albert S, Garanto A, Elmelik D, Qamar R, Lugtenberg D, van den Born LI, Collin RWJ, and Cremers FPM
- Subjects
- ATP-Binding Cassette Transporters biosynthesis, Adult, Female, Humans, Macular Degeneration genetics, Macular Degeneration metabolism, Male, RNA Precursors metabolism, Stargardt Disease, ATP-Binding Cassette Transporters genetics, Macular Degeneration congenital, RNA Precursors genetics, RNA Splice Sites, RNA Splicing
- Abstract
Stargardt disease is caused by variants in the ABCA4 gene, a significant part of which are noncanonical splice site (NCSS) variants. In case a gene of interest is not expressed in available somatic cells, small genomic fragments carrying potential disease-associated variants are tested for splice abnormalities using in vitro splice assays. We recently discovered that when using small minigenes lacking the proper genomic context, in vitro results do not correlate with splice defects observed in patient cells. We therefore devised a novel strategy in which a bacterial artificial chromosome was employed to generate midigenes, splice vectors of varying lengths (up to 11.7 kb) covering almost the entire ABCA4 gene. These midigenes were used to analyze the effect of all 44 reported and three novel NCSS variants on ABCA4 pre-mRNA splicing. Intriguingly, multi-exon skipping events were observed, as well as exon elongation and intron retention. The analysis of all reported NCSS variants in ABCA4 allowed us to reveal the nature of aberrant splicing events and to classify the severity of these mutations based on the residual fraction of wild-type mRNA. Our strategy to generate large overlapping splice vectors carrying multiple exons, creating a toolbox for robust and high-throughput analysis of splice variants, can be applied to all human genes., (© 2018 Sangermano et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2018
- Full Text
- View/download PDF
39. MIB2 variants altering NOTCH signalling result in left ventricle hypertrabeculation/non-compaction and are associated with Ménétrier-like gastropathy.
- Author
-
Piccolo P, Attanasio S, Secco I, Sangermano R, Strisciuglio C, Limongelli G, Miele E, Mutarelli M, Banfi S, Nigro V, Pons T, Valencia A, Zentilin L, Campione S, Nardone G, Lynnes TC, Celestino-Soper PB, Spoonamore KG, D'Armiento FP, Giacca M, Staiano A, Vatta M, Collesi C, and Brunetti-Pierri N
- Subjects
- Animals, Animals, Newborn, Cardiomyopathies etiology, Cardiomyopathies metabolism, Case-Control Studies, Cells, Cultured, Exome genetics, Female, Gastritis, Hypertrophic etiology, Gastritis, Hypertrophic metabolism, Gene Expression Regulation, Humans, Male, Myocytes, Cardiac cytology, Myocytes, Cardiac metabolism, Pedigree, Phenotype, Rats, Receptors, Notch genetics, Signal Transduction, Stomach Diseases etiology, Stomach Diseases metabolism, Ubiquitin-Protein Ligases metabolism, Ubiquitination, Ventricular Dysfunction, Left etiology, Ventricular Dysfunction, Left metabolism, Cardiomyopathies pathology, Gastritis, Hypertrophic pathology, Mutation, Missense genetics, Receptors, Notch metabolism, Stomach Diseases pathology, Ubiquitin-Protein Ligases genetics, Ventricular Dysfunction, Left pathology
- Abstract
We performed whole exome sequencing in individuals from a family with autosomal dominant gastropathy resembling Ménétrier disease, a premalignant gastric disorder with epithelial hyperplasia and enhanced EGFR signalling. Ménétrier disease is believed to be an acquired disorder, but its aetiology is unknown. In affected members, we found a missense p.V742G variant in MIB2, a gene regulating NOTCH signalling that has not been previously linked to human diseases. The variant segregated with the disease in the pedigree, affected a highly conserved amino acid residue, and was predicted to be deleterious although it was found with a low frequency in control individuals. The purified protein carrying the p.V742G variant showed reduced ubiquitination activity in vitro and white blood cells from affected individuals exhibited significant reductions of HES1 and NOTCH3 expression reflecting alteration of NOTCH signalling. Because mutations of MIB1, the homolog of MIB2, have been found in patients with left ventricle non-compaction (LVNC), we investigated members of our family with Ménétrier-like disease for this cardiac abnormality. Asymptomatic left ventricular hypertrabeculation, the mildest end of the LVNC spectrum, was detected in two members carrying the MIB2 variant. Finally, we identified an additional MIB2 variant (p.V984L) affecting protein stability in an unrelated isolated case with LVNC. Expression of both MIB2 variants affected NOTCH signalling, proliferation and apoptosis in primary rat cardiomyocytes.In conclusion, we report the first example of left ventricular hypertrabeculation/LVNC with germline MIB2 variants resulting in altered NOTCH signalling that might be associated with a gastropathy clinically overlapping with Ménétrier disease., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2017
- Full Text
- View/download PDF
40. Photoreceptor Progenitor mRNA Analysis Reveals Exon Skipping Resulting from the ABCA4 c.5461-10T→C Mutation in Stargardt Disease.
- Author
-
Sangermano R, Bax NM, Bauwens M, van den Born LI, De Baere E, Garanto A, Collin RW, Goercharn-Ramlal AS, den Engelsman-van Dijk AH, Rohrschneider K, Hoyng CB, Cremers FP, and Albert S
- Subjects
- Adult, DNA Mutational Analysis, Electroretinography, Female, Fibroblasts metabolism, Fluorescein Angiography, HEK293 Cells, Haplotypes, Humans, Induced Pluripotent Stem Cells metabolism, Macular Degeneration diagnosis, Macular Degeneration genetics, Macular Degeneration physiopathology, Male, Middle Aged, Polymorphism, Single Nucleotide, RNA Splice Sites genetics, Reverse Transcriptase Polymerase Chain Reaction, Stargardt Disease, Transfection, Visual Acuity physiology, Visual Fields physiology, Young Adult, ATP-Binding Cassette Transporters genetics, Alternative Splicing, Exons genetics, Macular Degeneration congenital, Photoreceptor Cells, Vertebrate physiology, RNA, Messenger genetics, Stem Cells physiology
- Abstract
Purpose: To elucidate the functional effect of the ABCA4 variant c.5461-10T→C, one of the most frequent variants associated with Stargardt disease (STGD1)., Design: Case series., Participants: Seventeen persons with STGD1 carrying ABCA4 variants and 1 control participant., Methods: Haplotype analysis of 4 homozygotes and 11 heterozygotes for c.5461-10T→C and sequence analysis of the ABCA4 gene for a homozygous proband. Fibroblasts were reprogrammed from 3 persons with STGD1 into induced pluripotent stem cells, which were differentiated into photoreceptor progenitor cells (PPCs). The effect of the c.5461-10T→C variant on RNA splicing by reverse-transcription polymerase chain reaction was analyzed using PPC mRNA. In vitro assays were performed with minigene constructs containing ABCA4 exon 39. We analyzed the natural history and ophthalmologic characteristics of 4 persons homozygous for c.5461-10T→C., Main Outcome Measures: Haplotype and rare variant data for ABCA4, RNA splice defects, age at diagnosis, visual acuity, fundus appearance, visual field, electroretinography (ERG) results, fluorescein angiography results, and fundus autofluorescence findings., Results: The frequent ABCA4 variant c.5461-10T→C has a subtle effect on splicing based on prediction programs. A founder haplotype containing c.5461-10T→C was found to span approximately 96 kb of ABCA4 and did not contain other rare sequence variants. Patient-derived PPCs showed skipping of exon 39 or exons 39 and 40 in the mRNA. HEK293T cell transduction with minigenes carrying exon 39 showed that the splice defects were the result of the c.5461-10T→C variant. All 4 subjects carrying the c.5461-10T→C variant in a homozygous state showed a young age of STGD1 onset, with low visual acuity at presentation and abnormal cone ERG results. All 4 demonstrated severe cone-rod dystrophy before 20 years of age and were legally blind by 25 years of age., Conclusions: The ABCA4 variant c.5461-10T→C is located on a founder haplotype lacking other disease-causing rare sequence variants. In vitro studies revealed that it leads to mRNA exon skipping and ABCA4 protein truncation. Given the severe phenotype in persons homozygous for this variant, we conclude that this variant results in the absence of ABCA4 activity., (Copyright © 2016 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
41. Mutations in MFSD8, encoding a lysosomal membrane protein, are associated with nonsyndromic autosomal recessive macular dystrophy.
- Author
-
Roosing S, van den Born LI, Sangermano R, Banfi S, Koenekoop RK, Zonneveld-Vrieling MN, Klaver CC, van Lith-Verhoeven JJ, Cremers FP, den Hollander AI, and Hoyng CB
- Subjects
- Adult, Aged, DNA Mutational Analysis, Electroretinography, Exome genetics, Female, Genome-Wide Association Study, Humans, Male, Middle Aged, Pedigree, Tomography, Optical Coherence, Visual Field Tests, Lysosomal Membrane Proteins genetics, Macular Degeneration genetics, Membrane Transport Proteins genetics, Mutation
- Abstract
Purpose: This study aimed to identify the genetic defects in 2 families with autosomal recessive macular dystrophy with central cone involvement., Design: Case series., Participants: Two families and a cohort of 244 individuals with various inherited maculopathies and cone disorders., Methods: Genome-wide linkage analysis and exome sequencing were performed in 1 large family with 5 affected individuals. In addition, exome sequencing was performed in the proband of a second family. Subsequent analysis of the identified mutations in 244 patients was performed by Sanger sequencing or restriction enzyme digestion. The medical history of individuals carrying the MFSD8 variants was reviewed and additional ophthalmic examinations were performed, including electroretinography (ERG), multifocal ERG (mfERG), perimetry, optical coherence tomography (OCT), fundus autofluorescence, and fundus photography., Main Outcome Measures: MFSD8 variants, age at diagnosis, visual acuity, fundus appearance, color vision defects, visual field, ERG, mfERG, fundus autofluorescence, and OCT findings., Results: Compound heterozygous variants in MFSD8, a gene encoding a lysosomal transmembrane protein, were identified in 2 families with macular dystrophy with a normal or subnormal ERG, but reduced mfERG. In both families, a heterozygous missense variant p.Glu336Gln was identified, which was predicted to have a mild effect on the protein. In the first family, a protein-truncating variant (p.Glu381*) was identified on the other allele, and in the second family, a variant (c.1102G>C) was identified that results in a splicing defect leading to skipping of exon 11 (p.Lys333Lysfs*3). The p.Glu336Gln allele was found to be significantly enriched in patients with maculopathies and cone disorders (6/488) compared with ethnically matched controls (35/18 682; P < 0.0001), suggesting that it may act as a genetic modifier., Conclusions: In this study, we identified variants in MFSD8 as a novel cause of nonsyndromic autosomal recessive macular dystrophy with central cone involvement. Affected individuals showed no neurologic features typical for variant late-infantile neuronal ceroid lipofuscinosis (vLINCL), a severe and devastating multisystem lysosomal storage disease previously associated with mutations in MFSD8. We propose a genotype-phenotype model in which a combination of a severe and a mild variant cause nonsyndromic macular dystrophy with central cone involvement, and 2 severe mutations cause vLINCL., (Copyright © 2015 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
42. Heterozygous deep-intronic variants and deletions in ABCA4 in persons with retinal dystrophies and one exonic ABCA4 variant.
- Author
-
Bax NM, Sangermano R, Roosing S, Thiadens AA, Hoefsloot LH, van den Born LI, Phan M, Klevering BJ, Westeneng-van Haaften C, Braun TA, Zonneveld-Vrieling MN, de Wijs I, Mutlu M, Stone EM, den Hollander AI, Klaver CC, Hoyng CB, and Cremers FP
- Subjects
- Exons, Female, Genetic Heterogeneity, Genetic Predisposition to Disease, High-Throughput Nucleotide Sequencing, Humans, Introns, Macular Degeneration genetics, Male, Pedigree, Sequence Analysis, DNA, Sequence Deletion, Stargardt Disease, ATP-Binding Cassette Transporters genetics, Genetic Association Studies methods, Macular Degeneration congenital, Retinitis Pigmentosa genetics
- Abstract
Variants in ABCA4 are responsible for autosomal-recessive Stargardt disease and cone-rod dystrophy. Sequence analysis of ABCA4 exons previously revealed one causative variant in each of 45 probands. To identify the "missing" variants in these cases, we performed multiplex ligation-dependent probe amplification-based deletion scanning of ABCA4. In addition, we sequenced the promoter region, fragments containing five deep-intronic splice variants, and 15 deep-intronic regions containing weak splice sites. Heterozygous deletions spanning ABCA4 exon 5 or exons 20-22 were found in two probands, heterozygous deep-intronic variants were identified in six probands, and a deep-intronic variant was found together with an exon 20-22 deletion in one proband. Based on ophthalmologic findings and characteristics of the identified exonic variants present in trans, the deep-intronic variants V1 and V4 were predicted to be relatively mild and severe, respectively. These findings are important for proper genetic counseling and for the development of variant-specific therapies., (© 2014 WILEY PERIODICALS, INC.)
- Published
- 2015
- Full Text
- View/download PDF
43. Gene decay. II. Analytic simulation of gene decay.
- Author
-
Rossi C and Sangermano R
- Subjects
- DNA metabolism, DNA Repair, Drug Stability, Mathematics, Models, Biological, Mutation, Probability, Time Factors, Genes
- Published
- 1972
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.