Gain-of-function mutations in RPA1 cause a syndrome with short telomeres and somatic genetic rescue
Richa Sharma
(1)
,
Sushree Sahoo
(1)
,
Masayoshi Honda
(2)
,
Sophie Granger
(2)
,
Charnise Goodings
(1)
,
Louis Sanchez
(3)
,
Axel Künstner
(4, 5)
,
Hauke Busch
(4, 5)
,
Fabian Beier
(6)
,
Shondra Pruett-Miller
(1)
,
Marcus Valentine
(1)
,
Alfonso Fernandez
(1)
,
Ti-Cheng Chang
(1)
,
Vincent Géli
(7, 8)
,
Dmitri Churikov
(7, 8)
,
Sandrine Hirschi
(9)
,
Victor Pastor
(1)
,
Melanie Boerries
(10, 11)
,
Melchior Lauten
(4)
,
Charikleia Kelaidi
(12)
,
Megan Cooper
(13)
,
Sarah Nicholas
,
Jill Rosenfeld
(14)
,
Sophia Polychronopoulou
(12)
,
Caroline Kannengiesser
(15, 16)
,
Carole Saintomé
(3, 17)
,
Charlotte Niemeyer
(18, 10)
,
Patrick Revy
(19)
,
Marc Wold
(2)
,
Maria Spies
(2)
,
Miriam Erlacher
(18, 10)
,
Stephane Coulon
(7, 8)
,
Marcin Wlodarski
(1, 10)
1
St Jude Children's Research Hospital
2 University of Iowa [Iowa City]
3 STRING - Structure et Instabilité des Génomes
4 Universität zu Lübeck = University of Lübeck [Lübeck]
5 UKSH - University Medical Center of Schleswig–Holstein = Universitätsklinikum Schleswig-Holstein
6 RWTH - Rheinisch-Westfälische Technische Hochschule Aachen University
7 CRCM - Centre de Recherche en Cancérologie de Marseille
8 IPC - Institut Paoli-Calmettes
9 CHU Strasbourg - Centre Hospitalier Universitaire [Strasbourg]
10 University of Freiburg [Freiburg]
11 Molecular Genetics of Breast Cancer, German Cancer Research Center (DKFZ)
12 Aghia Sophia Children's Hospital
13 Washington University School of Medicine in St. Louis
14 BCM - Baylor College of Medicine
15 AP-HP - Hôpital Bichat - Claude Bernard [Paris]
16 PHERE (UMR_S_1152 / U1152) - Physiopathologie et Epidémiologie des Maladies Respiratoires
17 UFR 927 - Sorbonne Université - UFR Sciences de la vie
18 DKFZ - German Cancer Research Center - Deutsches Krebsforschungszentrum [Heidelberg]
19 Imagine - U1163 - Imagine - Institut des maladies génétiques (IHU)
2 University of Iowa [Iowa City]
3 STRING - Structure et Instabilité des Génomes
4 Universität zu Lübeck = University of Lübeck [Lübeck]
5 UKSH - University Medical Center of Schleswig–Holstein = Universitätsklinikum Schleswig-Holstein
6 RWTH - Rheinisch-Westfälische Technische Hochschule Aachen University
7 CRCM - Centre de Recherche en Cancérologie de Marseille
8 IPC - Institut Paoli-Calmettes
9 CHU Strasbourg - Centre Hospitalier Universitaire [Strasbourg]
10 University of Freiburg [Freiburg]
11 Molecular Genetics of Breast Cancer, German Cancer Research Center (DKFZ)
12 Aghia Sophia Children's Hospital
13 Washington University School of Medicine in St. Louis
14 BCM - Baylor College of Medicine
15 AP-HP - Hôpital Bichat - Claude Bernard [Paris]
16 PHERE (UMR_S_1152 / U1152) - Physiopathologie et Epidémiologie des Maladies Respiratoires
17 UFR 927 - Sorbonne Université - UFR Sciences de la vie
18 DKFZ - German Cancer Research Center - Deutsches Krebsforschungszentrum [Heidelberg]
19 Imagine - U1163 - Imagine - Institut des maladies génétiques (IHU)
Masayoshi Honda
- Fonction : Auteur
- PersonId : 814853
- ORCID : 0000-0001-8920-6301
Charnise Goodings
- Fonction : Auteur
- PersonId : 814854
- ORCID : 0000-0001-8189-0989
Axel Künstner
- Fonction : Auteur
- PersonId : 779978
- ORCID : 0000-0003-0692-2105
Hauke Busch
- Fonction : Auteur
- PersonId : 814855
- ORCID : 0000-0003-4763-4521
Shondra Pruett-Miller
- Fonction : Auteur
- PersonId : 814856
- ORCID : 0000-0002-3793-585X
Vincent Géli
- Fonction : Auteur
- PersonId : 760479
- ORCID : 0000-0002-4103-7462
- IdRef : 033776601
Dmitri Churikov
- Fonction : Auteur
- PersonId : 776109
- ORCID : 0000-0003-1127-8852
Melanie Boerries
- Fonction : Auteur
- PersonId : 814857
- ORCID : 0000-0002-3670-0602
Melchior Lauten
- Fonction : Auteur
- PersonId : 763641
- ORCID : 0000-0003-4774-0279
Megan Cooper
- Fonction : Auteur
- PersonId : 814858
- ORCID : 0000-0002-5696-172X
Sarah Nicholas
- Fonction : Auteur
Jill Rosenfeld
- Fonction : Auteur
- PersonId : 794554
- ORCID : 0000-0001-5664-7987
Sophia Polychronopoulou
- Fonction : Auteur
- PersonId : 814859
- ORCID : 0000-0001-9716-9264
Carole Saintomé
- Fonction : Auteur
- PersonId : 757044
- ORCID : 0000-0002-7609-6913
- IdRef : 174901569
Charlotte Niemeyer
- Fonction : Auteur
- PersonId : 797656
- ORCID : 0000-0003-3856-7937
Patrick Revy
- Fonction : Auteur
- PersonId : 738943
- IdHAL : patrick-revy
- ORCID : 0000-0003-0758-8022
- IdRef : 154700126
Maria Spies
- Fonction : Auteur
- PersonId : 784875
- ORCID : 0000-0002-7375-8037
Stephane Coulon
- Fonction : Auteur
- PersonId : 179413
- IdHAL : stephane-coulon
- ORCID : 0000-0001-8090-914X
- IdRef : 089048393
Marcin Wlodarski
- Fonction : Auteur
- PersonId : 797655
- ORCID : 0000-0001-6638-9643
Résumé
Abstract Human telomere biology disorders (TBD)/short telomere syndromes (STS) are heterogeneous disorders caused by inherited loss-of-function mutations in telomere-associated genes. Here, we identify 3 germline heterozygous missense variants in the RPA1 gene in 4 unrelated probands presenting with short telomeres and varying clinical features of TBD/STS, including bone marrow failure, myelodysplastic syndrome, T- and B-cell lymphopenia, pulmonary fibrosis, or skin manifestations. All variants cluster to DNA-binding domain A of RPA1 protein. RPA1 is a single-strand DNA-binding protein required for DNA replication and repair and involved in telomere maintenance. We showed that RPA1E240K and RPA1V227A proteins exhibit increased binding to single-strand and telomeric DNA, implying a gain in DNA-binding function, whereas RPA1T270A has binding properties similar to wild-type protein. To study the mutational effect in a cellular system, CRISPR/Cas9 was used to knock-in the RPA1E240K mutation into healthy inducible pluripotent stem cells. This resulted in severe telomere shortening and impaired hematopoietic differentiation. Furthermore, in patients with RPA1E240K, we discovered somatic genetic rescue in hematopoietic cells due to an acquired truncating cis RPA1 mutation or a uniparental isodisomy 17p with loss of mutant allele, coinciding with stabilized blood counts. Using single-cell sequencing, the 2 somatic genetic rescue events were proven to be independently acquired in hematopoietic stem cells. In summary, we describe the first human disease caused by germline RPA1 variants in individuals with TBD/STS.