26 results on '"Hollizeck, S"'
Search Results
2. RET Solvent Front Mutations Mediate Acquired Resistance to Selective RET Inhibition in RET-Driven Malignancies
- Author
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Solomon, BJ, Tan, L, Lin, JJ, Wong, SQ, Hollizeck, S, Ebata, K, Tuch, BB, Yoda, S, Gainor, JF, Sequist, LV, Oxnard, GR, Gautschi, O, Drilon, A, Subbiah, V, Khoo, C, Zhu, EY, Nguyen, M, Henry, D, Condroski, KR, Kolakowski, GR, Gomez, E, Ballard, J, Metcalf, AT, Blake, JF, Dawson, S-J, Blosser, W, Stancato, LF, Brandhuber, BJ, Andrews, S, Robinson, BG, Rothenberg, SM, Solomon, BJ, Tan, L, Lin, JJ, Wong, SQ, Hollizeck, S, Ebata, K, Tuch, BB, Yoda, S, Gainor, JF, Sequist, LV, Oxnard, GR, Gautschi, O, Drilon, A, Subbiah, V, Khoo, C, Zhu, EY, Nguyen, M, Henry, D, Condroski, KR, Kolakowski, GR, Gomez, E, Ballard, J, Metcalf, AT, Blake, JF, Dawson, S-J, Blosser, W, Stancato, LF, Brandhuber, BJ, Andrews, S, Robinson, BG, and Rothenberg, SM
- Abstract
INTRODUCTION: Novel rearranged in transfection (RET)-specific tyrosine kinase inhibitors (TKIs) such as selpercatinib (LOXO-292) have shown unprecedented efficacy in tumors positive for RET fusions or mutations, notably RET fusion-positive NSCLC and RET-mutated medullary thyroid cancer (MTC). However, the mechanisms of resistance to these agents have not yet been described. METHODS: Analysis was performed of circulating tumor DNA and tissue in patients with RET fusion-positive NSCLC and RET-mutation positive MTC who developed disease progression after an initial response to selpercatinib. Acquired resistance was modeled preclinically using a CCDC6-RET fusion-positive NSCLC patient-derived xenograft. The inhibitory activity of anti-RET multikinase inhibitors and selective RET TKIs was evaluated in enzyme and cell-based assays. RESULTS: After a dramatic initial response to selpercatinib in a patient with KIF5B-RET NSCLC, analysis of circulating tumor DNA revealed emergence of RET G810R, G810S, and G810C mutations in the RET solvent front before the emergence of clinical resistance. Postmortem biopsy studies reported intratumor and intertumor heterogeneity with distinct disease subclones containing G810S, G810R, and G810C mutations in multiple disease sites indicative of convergent evolution on the G810 residue resulting in a common mechanism of resistance. Acquired mutations in RET G810 were identified in tumor tissue from a second patient with CCDC6-RET fusion-positive NSCLC and in plasma from patients with additional RET fusion-positive NSCLC and RET-mutant MTC progressing on an ongoing phase 1 and 2 trial of selpercatinib. Preclinical studies reported the presence of RET G810R mutations in a CCDC6-RET patient-derived xenograft (from a patient with NSCLC) model of acquired resistance to selpercatinib. Structural modeling predicted that these mutations sterically hinder the binding of selpercatinib, and in vitro assays confirmed loss of activity for both anti-RET mul
- Published
- 2020
3. Human FCHO1 deficiency reveals role for clathrin-mediated endocytosis in development and function of T cells
- Author
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Lyszkiewicz, M, Zietara, N, Frey, L, Pannicke, U, Stern, M, Liu, Y, Fan, Y, Puchalka, J, Hollizeck, S, Somekh, I, Rohlfs, M, Yilmaz, T, Unal, E, Karakukcu, M, Patiroglu, T, Kellerer, C, Karasu, E, Sykora, K-W, Lev, A, Simon, A, Somech, R, Roesler, J, Hoenig, M, Keppler, OT, Schwarz, K, Klein, C, Lyszkiewicz, M, Zietara, N, Frey, L, Pannicke, U, Stern, M, Liu, Y, Fan, Y, Puchalka, J, Hollizeck, S, Somekh, I, Rohlfs, M, Yilmaz, T, Unal, E, Karakukcu, M, Patiroglu, T, Kellerer, C, Karasu, E, Sykora, K-W, Lev, A, Simon, A, Somech, R, Roesler, J, Hoenig, M, Keppler, OT, Schwarz, K, and Klein, C
- Abstract
Clathrin-mediated endocytosis (CME) is critical for internalisation of molecules across cell membranes. The FCH domain only 1 (FCHO1) protein is key molecule involved in the early stages of CME formation. The consequences of mutations in FCHO1 in humans were unknown. We identify ten unrelated patients with variable T and B cell lymphopenia, who are homozygous for six distinct mutations in FCHO1. We demonstrate that these mutations either lead to mislocalisation of the protein or prevent its interaction with binding partners. Live-cell imaging of cells expressing mutant variants of FCHO1 provide evidence of impaired formation of clathrin coated pits (CCP). Patient T cells are unresponsive to T cell receptor (TCR) triggering. Internalisation of the TCR receptor is severely perturbed in FCHO1-deficient Jurkat T cells but can be rescued by expression of wild-type FCHO1. Thus, we discovered a previously unrecognised critical role of FCHO1 and CME during T-cell development and function in humans.
- Published
- 2020
4. Human FCHO1 deficiency reveals role for clathrin-mediated endocytosis in development and function of T cells (vol 11, 1031, 2020)
- Author
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Lyszkiewicz, M, Zietara, N, Frey, L, Pannicke, U, Stern, M, Liu, Y, Fan, Y, Puchalka, J, Hollizeck, S, Somekh, I, Rohlfs, M, Yilmaz, T, Unal, E, Karakukcu, M, Patiroglu, T, Kellerer, C, Karasu, E, Sykora, K-W, Lev, A, Simon, A, Somech, R, Roesler, J, Hoenig, M, Keppler, OT, Schwarz, K, Klein, C, Lyszkiewicz, M, Zietara, N, Frey, L, Pannicke, U, Stern, M, Liu, Y, Fan, Y, Puchalka, J, Hollizeck, S, Somekh, I, Rohlfs, M, Yilmaz, T, Unal, E, Karakukcu, M, Patiroglu, T, Kellerer, C, Karasu, E, Sykora, K-W, Lev, A, Simon, A, Somech, R, Roesler, J, Hoenig, M, Keppler, OT, Schwarz, K, and Klein, C
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
- Published
- 2020
5. Proteome Analysis of Human Neutrophil Granulocytes From Patients With Monogenic Disease Using Data-independent Acquisition
- Author
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Grabowski, P, Hesse, S, Hollizeck, S, Rohlfs, M, Behrends, U, Sherkat, R, Tamary, H, Unal, E, Somech, R, Patiroglu, T, Canzar, S, Ten Bosch, JVDW, Klein, C, Rappsilber, J, Grabowski, P, Hesse, S, Hollizeck, S, Rohlfs, M, Behrends, U, Sherkat, R, Tamary, H, Unal, E, Somech, R, Patiroglu, T, Canzar, S, Ten Bosch, JVDW, Klein, C, and Rappsilber, J
- Abstract
Neutrophil granulocytes are critical mediators of innate immunity and tissue regeneration. Rare diseases of neutrophil granulocytes may affect their differentiation and/or functions. However, there are very few validated diagnostic tests assessing the functions of neutrophil granulocytes in these diseases. Here, we set out to probe omics analysis as a novel diagnostic platform for patients with defective differentiation and function of neutrophil granulocytes. We analyzed highly purified neutrophil granulocytes from 68 healthy individuals and 16 patients with rare monogenic diseases. Cells were isolated from fresh venous blood (purity >99%) and used to create a spectral library covering almost 8000 proteins using strong cation exchange fractionation. Patient neutrophil samples were then analyzed by data-independent acquisition proteomics, quantifying 4154 proteins in each sample. Neutrophils with mutations in the neutrophil elastase gene ELANE showed large proteome changes that suggest these mutations may affect maturation of neutrophil granulocytes and initiate misfolded protein response and cellular stress mechanisms. In contrast, only few proteins changed in patients with leukocyte adhesion deficiency (LAD) and chronic granulomatous disease (CGD). Strikingly, neutrophil transcriptome analysis showed no correlation with its proteome. In case of two patients with undetermined genetic causes, proteome analysis guided the targeted genetic diagnostics and uncovered the underlying genomic mutations. Data-independent acquisition proteomics may help to define novel pathomechanisms in neutrophil diseases and provide a clinically useful diagnostic dimension.
- Published
- 2019
6. Unravelling mutational signatures with plasma circulating tumour DNA.
- Author
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Hollizeck S, Wang N, Wong SQ, Litchfield C, Guinto J, Ftouni S, Rebello R, Kanwal S, Dong R, Grimmond S, Sandhu S, Mileshkin L, Tothill RW, Chandrananda D, and Dawson SJ
- Subjects
- Humans, Liquid Biopsy methods, Whole Genome Sequencing methods, DNA Mutational Analysis methods, Biomarkers, Tumor genetics, Biomarkers, Tumor blood, DNA, Neoplasm genetics, DNA, Neoplasm blood, Circulating Tumor DNA genetics, Circulating Tumor DNA blood, Mutation, Neoplasms genetics, Neoplasms blood
- Abstract
The use of circulating tumour DNA (ctDNA) to profile mutational signatures represents a non-invasive opportunity for understanding cancer mutational processes. Here we present MisMatchFinder, a liquid biopsy approach for mutational signature detection using low-coverage whole-genome sequencing of ctDNA. Through analysis of 375 plasma samples across 9 cancers, we demonstrate that MisMatchFinder accurately infers single-base and doublet-base substitutions, as well as insertions and deletions to enhance the detection of ctDNA and clinically relevant mutational signatures., Competing Interests: Competing interests The authors declare the following competing interests: S.-J.D. is an advisory board member for Adela. The remaining authors declare no competing interests., (© 2024. The Author(s).)
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- 2024
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7. Prospective cohort study of genomic newborn screening: BabyScreen+ pilot study protocol.
- Author
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Lunke S, Bouffler SE, Downie L, Caruana J, Amor DJ, Archibald A, Bombard Y, Christodoulou J, Clausen M, De Fazio P, Greaves RF, Hollizeck S, Kanga-Parabia A, Lang N, Lynch F, Peters R, Sadedin S, Tutty E, Eggers S, Lee C, Wall M, Yeung A, Gaff C, Gyngell C, Vears DF, Best S, Goranitis I, and Stark Z
- Subjects
- Child, Humans, Infant, Newborn, Pilot Projects, Prospective Studies, Victoria, Neonatal Screening, Genomics
- Abstract
Introduction: Newborn bloodspot screening (NBS) is a highly successful public health programme that uses biochemical and other assays to screen for severe but treatable childhood-onset conditions. Introducing genomic sequencing into NBS programmes increases the range of detectable conditions but raises practical and ethical issues. Evidence from prospectively ascertained cohorts is required to guide policy and future implementation. This study aims to develop, implement and evaluate a genomic NBS (gNBS) pilot programme., Methods and Analysis: The BabyScreen+ study will pilot gNBS in three phases. In the preimplementation phase, study materials, including education resources, decision support and data collection tools, will be designed. Focus groups and key informant interviews will also be undertaken to inform delivery of the study and future gNBS programmes. During the implementation phase, we will prospectively recruit birth parents in Victoria, Australia, to screen 1000 newborns for over 600 severe, treatable, childhood-onset conditions. Clinically accredited whole genome sequencing will be performed following standard NBS using the same sample. High chance results will be returned by genetic healthcare professionals, with follow-on genetic and other confirmatory testing and referral to specialist services as required. The postimplementation phase will evaluate the feasibility of gNBS as the primary aim, and assess ethical, implementation, psychosocial and health economic factors to inform future service delivery., Ethics and Dissemination: This project received ethics approval from the Royal Children's Hospital Melbourne Research Ethics Committee: HREC/91500/RCHM-2023, HREC/90929/RCHM-2022 and HREC/91392/RCHM-2022. Findings will be disseminated to policy-makers, and through peer-reviewed journals and conferences., Competing Interests: Competing interests: YB and MC are cofounders of the Genetics Adviser. The other authors declare no relevant disclosures., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
- Published
- 2024
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8. Valosin-containing protein-regulated endoplasmic reticulum stress causes NOD2-dependent inflammatory responses.
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Ghalandary M, Li Y, Fröhlich T, Magg T, Liu Y, Rohlfs M, Hollizeck S, Conca R, Schwerd T, Uhlig HH, Bufler P, Koletzko S, Muise AM, Snapper SB, Hauck F, Klein C, and Kotlarz D
- Subjects
- Acetylmuramyl-Alanyl-Isoglutamine pharmacology, Humans, Nod2 Signaling Adaptor Protein genetics, Nod2 Signaling Adaptor Protein metabolism, Valosin Containing Protein genetics, Valosin Containing Protein metabolism, Endoplasmic Reticulum Stress, Interleukin-8 metabolism
- Abstract
NOD2 polymorphisms may affect sensing of the bacterial muramyl dipeptide (MDP) and trigger perturbed inflammatory responses. Genetic screening of a patient with immunodeficiency and enteropathy revealed a rare homozygous missense mutation in the first CARD domain of NOD2 (ENST00000300589; c.160G > A, p.E54K). Biochemical assays confirmed impaired NOD2-dependent signaling and proinflammatory cytokine production in patient's cells and heterologous cellular models with overexpression of the NOD2 mutant. Immunoprecipitation-coupled mass spectrometry unveiled the ATPase valosin-containing protein (VCP) as novel interaction partner of wildtype NOD2, while the binding to the NOD2 variant p.E54K was abrogated. Knockdown of VCP in coloncarcinoma cells led to impaired NF-κB activity and IL8 expression upon MDP stimulation. In contrast, tunicamycin-induced ER stress resulted in increased IL8, CXCL1, and CXCL2 production in cells with knockdown of VCP, while enhanced expression of these proinflammatory molecules was abolished upon knockout of NOD2. Taken together, these data suggest that VCP-mediated inflammatory responses upon ER stress are NOD2-dependent., (© 2022. The Author(s).)
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- 2022
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9. Non-genetic determinants of malignant clonal fitness at single-cell resolution.
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Fennell KA, Vassiliadis D, Lam EYN, Martelotto LG, Balic JJ, Hollizeck S, Weber TS, Semple T, Wang Q, Miles DC, MacPherson L, Chan YC, Guirguis AA, Kats LM, Wong ES, Dawson SJ, Naik SH, and Dawson MA
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- Animals, Cell Line, Cell Lineage drug effects, Clone Cells drug effects, Clone Cells metabolism, Female, Humans, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myeloid, Acute genetics, Mice, Mice, Inbred C57BL, Secretory Leukocyte Peptidase Inhibitor metabolism, Cell Competition drug effects, Clone Cells pathology, Leukemia, Myeloid, Acute pathology, Single-Cell Analysis
- Abstract
All cancers emerge after a period of clonal selection and subsequent clonal expansion. Although the evolutionary principles imparted by genetic intratumour heterogeneity are becoming increasingly clear
1 , little is known about the non-genetic mechanisms that contribute to intratumour heterogeneity and malignant clonal fitness2 . Here, using single-cell profiling and lineage tracing (SPLINTR)-an expressed barcoding strategy-we trace isogenic clones in three clinically relevant mouse models of acute myeloid leukaemia. We find that malignant clonal dominance is a cell-intrinsic and heritable property that is facilitated by the repression of antigen presentation and increased expression of the secretory leukocyte peptidase inhibitor gene (Slpi), which we genetically validate as a regulator of acute myeloid leukaemia. Increased transcriptional heterogeneity is a feature that enables clonal fitness in diverse tissues and immune microenvironments and in the context of clonal competition between genetically distinct clones. Similar to haematopoietic stem cells3 , leukaemia stem cells (LSCs) display heritable clone-intrinsic properties of high, and low clonal output that contribute to the overall tumour mass. We demonstrate that LSC clonal output dictates sensitivity to chemotherapy and, although high- and low-output clones adapt differently to therapeutic pressure, they coordinately emerge from minimal residual disease with increased expression of the LSC program. Together, these data provide fundamental insights into the non-genetic transcriptional processes that underpin malignant clonal fitness and may inform future therapeutic strategies., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2022
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10. NOX1 Regulates Collective and Planktonic Cell Migration: Insights From Patients With Pediatric-Onset IBD and NOX1 Deficiency.
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Khoshnevisan R, Anderson M, Babcock S, Anderson S, Illig D, Marquardt B, Sherkat R, Schröder K, Moll F, Hollizeck S, Rohlfs M, Walz C, Adibi P, Rezaei A, Andalib A, Koletzko S, Muise AM, Snapper SB, Klein C, Thiagarajah JR, and Kotlarz D
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- Adolescent, Adult, Cell Line, Child, Cytoskeletal Proteins metabolism, Female, Humans, Male, Mutation, Young Adult, Cell Movement genetics, Inflammatory Bowel Diseases genetics, Intestinal Mucosa cytology, NADPH Oxidase 1 deficiency, Wound Healing genetics
- Abstract
Background: Genetic defects of pediatric-onset inflammatory bowel disease (IBD) provide critical insights into molecular factors controlling intestinal homeostasis. NOX1 has been recently recognized as a major source of reactive oxygen species (ROS) in human colonic epithelial cells. Here we assessed the functional consequences of human NOX1 deficiency with respect to wound healing and epithelial migration by studying pediatric IBD patients presenting with a stop-gain mutation in NOX1., Methods: Functional characterization of the NOX1 variant included ROS generation, wound healing, 2-dimensional collective chemotactic migration, single-cell planktonic migration in heterologous cell lines, and RNA scope and immunohistochemistry of paraffin-embedded patient tissue samples., Results: Using exome sequencing, we identified a stop-gain mutation in NOX1 (c.160C>T, p.54R>*) in patients with pediatric-onset IBD. Our studies confirmed that loss-of-function of NOX1 causes abrogated ROS activity, but they also provided novel mechanistic insights into human NOX1 deficiency. Cells that were NOX1-mutant showed impaired wound healing and attenuated 2-dimensional collective chemotactic migration. High-resolution microscopy of the migrating cell edge revealed a reduced density of filopodial protrusions with altered focal adhesions in NOX1-deficient cells, accompanied by reduced phosphorylation of p190A. Assessment of single-cell planktonic migration toward an epidermal growth factor gradient showed that NOX1 deficiency is associated with altered migration dynamics with loss of directionality and altered cell-cell interactions., Conclusions: Our studies on pediatric-onset IBD patients with a rare sequence variant in NOX1 highlight that human NOX1 is involved in regulating wound healing by altering epithelial cytoskeletal dynamics at the leading edge and directing cell migration., (© 2020 Crohn’s & Colitis Foundation. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2020
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11. Author Correction: Human FCHO1 deficiency reveals role for clathrin-mediated endocytosis in development and function of T cells.
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Łyszkiewicz M, Ziętara N, Frey L, Pannicke U, Stern M, Liu Y, Fan Y, Puchałka J, Hollizeck S, Somekh I, Rohlfs M, Yilmaz T, Ünal E, Karakukcu M, Patiroğlu T, Kellerer C, Karasu E, Sykora KW, Lev A, Simon A, Somech R, Roesler J, Hoenig M, Keppler OT, Schwarz K, and Klein C
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
- Published
- 2020
- Full Text
- View/download PDF
12. Lymphocytic interstitial pneumonia and follicular bronchiolitis in children: A registry-based case series.
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Prenzel F, Harfst J, Schwerk N, Ahrens F, Rietschel E, Schmitt-Grohé S, Rubak SML, Poplawska K, Baden W, Vogel M, Hollizeck S, Ley-Zaporozhan J, Brasch F, Reu S, and Griese M
- Subjects
- Adolescent, Age of Onset, Biopsy, Bronchitis diagnosis, Bronchitis drug therapy, Bronchitis pathology, Child, Child, Preschool, Chronic Disease, Cough etiology, Diagnosis, Differential, Dyspnea etiology, Female, Genetic Testing, Humans, Infant, Lung diagnostic imaging, Lung pathology, Lung Diseases diagnosis, Lung Diseases, Interstitial diagnosis, Lung Diseases, Interstitial drug therapy, Lung Diseases, Interstitial pathology, Male, Registries, Respiratory Sounds etiology, Respiratory Tract Infections etiology, Retrospective Studies, Tomography, X-Ray Computed, Bronchitis complications, Lung Diseases, Interstitial complications
- Abstract
Objectives: Pediatric lymphocytic interstitial pneumonia (LIP) and follicular bronchiolitis (FB) are poorly characterized lymphoproliferative disorders. We present and quantify demographics, radiological and histopathologic patterns, treatments and their responses, and outcomes in non-HIV-infected children with LIP and FB., Methods: This structured registry-based study included a retrospective chart review, blinded analysis of imaging studies and lung biopsies, genetic testing, and evaluation of treatments and outcomes., Results: Of the 13 patients (eight females) studied, eight had FB, four had combined LIP/FB, and one had isolated LIP; diagnoses were highly concordant between the pathologists. Most patients became symptomatic during the first 2 years of life, with a mean lag time to diagnosis of 4 years. The most common symptoms were coughing and respiratory infections (11 out of 13 each), dyspnea (10 out of 13), and wheezing (eight out of 13). Autoantibodies were found in eight out of 13 patients. In three patients, disease-causing mutations in the COPA gene were identified. CT revealed hilar lymphadenopathy (five out of 12), ground-glass opacity (eight out of 12), consolidation (five out of 12), and cysts (four out of 13). Systemic steroids as intravenous pulses (11 out of 13) or oral intake (10 out of 13) were the main treatments and showed high response rates of 100% and 90%, respectively. Within the mean observation period of 68 months, all children had chronic courses, eight out of 13 had severe diseases, two died, and one worsened., Conclusions: Children with LIP/FB have chronic diseases that occurred in early childhood and were commonly associated with immune dysregulation as well as high morbidity and mortality. Early diagnosis and treatment may be crucial to improve the outcome., (© 2020 Wiley Periodicals, Inc.)
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- 2020
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13. RET Solvent Front Mutations Mediate Acquired Resistance to Selective RET Inhibition in RET-Driven Malignancies.
- Author
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Solomon BJ, Tan L, Lin JJ, Wong SQ, Hollizeck S, Ebata K, Tuch BB, Yoda S, Gainor JF, Sequist LV, Oxnard GR, Gautschi O, Drilon A, Subbiah V, Khoo C, Zhu EY, Nguyen M, Henry D, Condroski KR, Kolakowski GR, Gomez E, Ballard J, Metcalf AT, Blake JF, Dawson SJ, Blosser W, Stancato LF, Brandhuber BJ, Andrews S, Robinson BG, and Rothenberg SM
- Subjects
- Humans, Mutation, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Pyrazoles, Pyridines, Solvents, Transfection, Lung Neoplasms drug therapy, Lung Neoplasms genetics, Proto-Oncogene Proteins c-ret genetics
- Abstract
Introduction: Novel rearranged in transfection (RET)-specific tyrosine kinase inhibitors (TKIs) such as selpercatinib (LOXO-292) have shown unprecedented efficacy in tumors positive for RET fusions or mutations, notably RET fusion-positive NSCLC and RET-mutated medullary thyroid cancer (MTC). However, the mechanisms of resistance to these agents have not yet been described., Methods: Analysis was performed of circulating tumor DNA and tissue in patients with RET fusion-positive NSCLC and RET-mutation positive MTC who developed disease progression after an initial response to selpercatinib. Acquired resistance was modeled preclinically using a CCDC6-RET fusion-positive NSCLC patient-derived xenograft. The inhibitory activity of anti-RET multikinase inhibitors and selective RET TKIs was evaluated in enzyme and cell-based assays., Results: After a dramatic initial response to selpercatinib in a patient with KIF5B-RET NSCLC, analysis of circulating tumor DNA revealed emergence of RET G810R, G810S, and G810C mutations in the RET solvent front before the emergence of clinical resistance. Postmortem biopsy studies reported intratumor and intertumor heterogeneity with distinct disease subclones containing G810S, G810R, and G810C mutations in multiple disease sites indicative of convergent evolution on the G810 residue resulting in a common mechanism of resistance. Acquired mutations in RET G810 were identified in tumor tissue from a second patient with CCDC6-RET fusion-positive NSCLC and in plasma from patients with additional RET fusion-positive NSCLC and RET-mutant MTC progressing on an ongoing phase 1 and 2 trial of selpercatinib. Preclinical studies reported the presence of RET G810R mutations in a CCDC6-RET patient-derived xenograft (from a patient with NSCLC) model of acquired resistance to selpercatinib. Structural modeling predicted that these mutations sterically hinder the binding of selpercatinib, and in vitro assays confirmed loss of activity for both anti-RET multikinase inhibitors and selective RET TKIs., Conclusions: RET G810 solvent front mutations represent the first described recurrent mechanism of resistance to selective RET inhibition with selpercatinib. Development of potent inhibitor of these mutations and maintaining activity against RET gatekeeper mutations could be an effective strategy to target resistance to selective RET inhibitors., (Copyright © 2020 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.)
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- 2020
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14. Human FCHO1 deficiency reveals role for clathrin-mediated endocytosis in development and function of T cells.
- Author
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Łyszkiewicz M, Ziętara N, Frey L, Pannicke U, Stern M, Liu Y, Fan Y, Puchałka J, Hollizeck S, Somekh I, Rohlfs M, Yilmaz T, Ünal E, Karakukcu M, Patiroğlu T, Kellerer C, Karasu E, Sykora KW, Lev A, Simon A, Somech R, Roesler J, Hoenig M, Keppler OT, Schwarz K, and Klein C
- Subjects
- Animals, CD4-Positive T-Lymphocytes pathology, Cell Differentiation, Cells, Cultured, Female, HIV Infections genetics, HIV-1 pathogenicity, Humans, Jurkat Cells, Lymphopenia pathology, Male, Membrane Proteins chemistry, Membrane Proteins genetics, Membrane Proteins metabolism, Mice, Pedigree, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes virology, Endocytosis physiology, Loss of Function Mutation, Lymphopenia genetics, Membrane Proteins deficiency, T-Lymphocytes physiology
- Abstract
Clathrin-mediated endocytosis (CME) is critical for internalisation of molecules across cell membranes. The FCH domain only 1 (FCHO1) protein is key molecule involved in the early stages of CME formation. The consequences of mutations in FCHO1 in humans were unknown. We identify ten unrelated patients with variable T and B cell lymphopenia, who are homozygous for six distinct mutations in FCHO1. We demonstrate that these mutations either lead to mislocalisation of the protein or prevent its interaction with binding partners. Live-cell imaging of cells expressing mutant variants of FCHO1 provide evidence of impaired formation of clathrin coated pits (CCP). Patient T cells are unresponsive to T cell receptor (TCR) triggering. Internalisation of the TCR receptor is severely perturbed in FCHO1-deficient Jurkat T cells but can be rescued by expression of wild-type FCHO1. Thus, we discovered a previously unrecognised critical role of FCHO1 and CME during T-cell development and function in humans.
- Published
- 2020
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15. Impact on Clinical Decision Making of Next-Generation Sequencing in Pediatric Epilepsy in a Tertiary Epilepsy Referral Center.
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Hoelz H, Herdl C, Gerstl L, Tacke M, Vill K, von Stuelpnagel C, Rost I, Hoertnagel K, Abicht A, Hollizeck S, Larsen LHG, and Borggraefe I
- Subjects
- Adolescent, Adult, Child, Child, Preschool, Electroencephalography methods, Female, Genetic Testing, Humans, Infant, Male, Prospective Studies, Referral and Consultation, Tertiary Care Centers, Young Adult, Clinical Decision-Making, Epilepsy genetics, Epilepsy physiopathology, High-Throughput Nucleotide Sequencing methods
- Abstract
Background . Next-generation sequencing (NGS) describes new powerful techniques of nucleic acid analysis, which allow not only disease gene identification diagnostics but also applications for transcriptome/methylation analysis and meta-genomics. NGS helps identify many monogenic epilepsy syndromes. Pediatric epilepsy patients can be tested using NGS epilepsy panels to diagnose them, thereby influencing treatment choices. The primary objective of this study was to evaluate the impact of genetic testing on clinical decision making in pediatric epilepsy patients. Methods . We completed a single-center retrospective cohort study of 91 patients (43 male) aged 19 years or less undergoing NGS with epilepsy panels differing in size ranging from 5 to 434 genes from October 2013 to September 2017. Results . During a mean time of 3.6 years between symptom onset and genetic testing, subjects most frequently showed epileptic encephalopathy (40%), focal epilepsy (33%), and generalized epilepsy (18%). In 16 patients (18% of the study population), "pathogenic" or "likely pathogenic" results according to ACMG criteria were found. Ten of the 16 patients (63%) experienced changes in clinical management regarding their medication and avoidance of further diagnostic evaluation, that is, presurgical evaluation. Conclusion . NGS epilepsy panels contribute to the diagnosis of pediatric epilepsy patients and may change their clinical management with regard to both preventing unnecessary and potentially harmful diagnostic procedures and management. Thus, the present data support the early implementation in order to adopt clinical management in selected cases and prevent further invasive investigations. Given the relatively small sample size and heterogeneous panels a larger prospective study with more homogeneous panels would be helpful to further determine the impact of NGS on clinical decision making.
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- 2020
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16. CARMIL2 Deficiency Presenting as Very Early Onset Inflammatory Bowel Disease.
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Magg T, Shcherbina A, Arslan D, Desai MM, Wall S, Mitsialis V, Conca R, Unal E, Karacabey N, Mukhina A, Rodina Y, Taur PD, Illig D, Marquardt B, Hollizeck S, Jeske T, Gothe F, Schober T, Rohlfs M, Koletzko S, Lurz E, Muise AM, Snapper SB, Hauck F, Klein C, and Kotlarz D
- Subjects
- Age of Onset, Child, Child, Preschool, Female, Genetic Predisposition to Disease, Humans, Lymphocytes immunology, Male, Mutation, Phenotype, Exome Sequencing, Inflammatory Bowel Diseases genetics, Inflammatory Bowel Diseases immunology, Microfilament Proteins deficiency
- Abstract
Background: Children with very early onset inflammatory bowel diseases (VEO-IBD) often have a refractory and severe disease course. A significant number of described VEO-IBD-causing monogenic disorders can be attributed to defects in immune-related genes. The diagnosis of the underlying primary immunodeficiency (PID) often has critical implications for the treatment of patients with IBD-like phenotypes., Methods: To identify the molecular etiology in 5 patients from 3 unrelated kindred with IBD-like symptoms, we conducted whole exome sequencing. Immune workup confirmed an underlying PID., Results: Whole exome sequencing revealed 3 novel CARMIL2 loss-of-function mutations in our patients. Immunophenotyping of peripheral blood mononuclear cells showed reduction of regulatory and effector memory T cells and impaired B cell class switching. The T cell proliferation and activation assays confirmed defective responses to CD28 costimulation, consistent with CARMIL2 deficiency., Conclusion: Our study highlights that human CARMIL2 deficiency can manifest with IBD-like symptoms. This example illustrates that early diagnosis of underlying PID is crucial for the treatment and prognosis of children with VEO-IBD., (© 2019 Crohn’s & Colitis Foundation. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2019
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17. An Evolutionarily Conserved Function of Polycomb Silences the MHC Class I Antigen Presentation Pathway and Enables Immune Evasion in Cancer.
- Author
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Burr ML, Sparbier CE, Chan KL, Chan YC, Kersbergen A, Lam EYN, Azidis-Yates E, Vassiliadis D, Bell CC, Gilan O, Jackson S, Tan L, Wong SQ, Hollizeck S, Michalak EM, Siddle HV, McCabe MT, Prinjha RK, Guerra GR, Solomon BJ, Sandhu S, Dawson SJ, Beavis PA, Tothill RW, Cullinane C, Lehner PJ, Sutherland KD, and Dawson MA
- Subjects
- Animals, Antigen Presentation drug effects, Antigen Presentation immunology, Antineoplastic Agents, Immunological pharmacology, Antineoplastic Agents, Immunological therapeutic use, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Cell Line, Tumor, DNA Methylation immunology, Down-Regulation drug effects, Down-Regulation genetics, Down-Regulation immunology, Drug Resistance, Neoplasm genetics, Epigenetic Repression drug effects, Epigenetic Repression immunology, Female, Gene Expression Regulation, Neoplastic drug effects, Histocompatibility Antigens Class I immunology, Histocompatibility Antigens Class I metabolism, Histone Code drug effects, Humans, Mice, Middle Aged, Neoplasms drug therapy, Neoplasms genetics, Neoplasms pathology, Polycomb Repressive Complex 2 antagonists & inhibitors, T-Lymphocytes immunology, Tumor Escape drug effects, Xenograft Model Antitumor Assays, Antineoplastic Combined Chemotherapy Protocols pharmacology, Gene Expression Regulation, Neoplastic immunology, Histocompatibility Antigens Class I genetics, Neoplasms immunology, Polycomb Repressive Complex 2 metabolism, Tumor Escape genetics
- Abstract
Loss of MHC class I (MHC-I) antigen presentation in cancer cells can elicit immunotherapy resistance. A genome-wide CRISPR/Cas9 screen identified an evolutionarily conserved function of polycomb repressive complex 2 (PRC2) that mediates coordinated transcriptional silencing of the MHC-I antigen processing pathway (MHC-I APP), promoting evasion of T cell-mediated immunity. MHC-I APP gene promoters in MHC-I low cancers harbor bivalent activating H3K4me3 and repressive H3K27me3 histone modifications, silencing basal MHC-I expression and restricting cytokine-induced upregulation. Bivalent chromatin at MHC-I APP genes is a normal developmental process active in embryonic stem cells and maintained during neural progenitor differentiation. This physiological MHC-I silencing highlights a conserved mechanism by which cancers arising from these primitive tissues exploit PRC2 activity to enable immune evasion., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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18. Genetic Deficiency and Biochemical Inhibition of ITK Affect Human Th17, Treg, and Innate Lymphoid Cells.
- Author
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Eken A, Cansever M, Somekh I, Mizoguchi Y, Zietara N, Okus FZ, Erdem S, Canatan H, Akyol S, Ozcan A, Karakukcu M, Hollizeck S, Rohlfs M, Unal E, Klein C, and Patiroglu T
- Subjects
- Animals, Apoptosis, Biomarkers, Cell Proliferation, Child, Preschool, Consanguinity, Cytokines metabolism, DNA Mutational Analysis, Female, Flow Cytometry, Forkhead Transcription Factors metabolism, High-Throughput Nucleotide Sequencing, Humans, Mice, Pedigree, Protein-Tyrosine Kinases metabolism, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets metabolism, T-Lymphocytes, Regulatory immunology, Th17 Cells immunology, Genetic Association Studies, Genetic Predisposition to Disease, Immunity, Innate, Protein-Tyrosine Kinases antagonists & inhibitors, Protein-Tyrosine Kinases deficiency, T-Lymphocytes, Regulatory metabolism, Th17 Cells metabolism
- Abstract
Purpose: Interleukin-2-inducible T cell kinase (ITK) is an important mediator of T cell receptor signaling. Loss of function mutations in ITK results in hypogammaglobulinemia and CD4+ T cell loss in humans, and the patients often present with EBV-associated B cell lymphoproliferative syndrome. Itk-deficient mice show loss of T cell naivety, impaired cytolytic activity of CD8+ T cells, and defects in CD4+ T cell lineage choice decisions. In mice, Itk mutations were shown to affect Th17-Treg lineage choice in favor of the latter. In this study, we explored whether human ITK reciprocally regulates Th17-Treg balance as its murine ortholog., Methods: Whole Exome Sequencing was used to identify the mutation. ITK-deficient peripheral blood lymphocytes were characterized by FACSAria III-based flow cytometric assays with respect to proliferation, apoptosis, cytokine production, and innate lymphoid cell (ILC) frequency. Sorted T cells from healthy donors were exposed to ibrutinib, an irreversible ITK inhibitor, to assess ITK's contribution to Th17 and Treg cell generation and functions., Results: In this study, we report a child with a novel ITK mutation who showed impaired CD3/CD28 induced proliferation in T cells. ITK-mutant cells were more apoptotic irrespective of TCR activation. More importantly, T cells produced less Th17-associated cytokines IL-17A, IL-22, and GM-CSF. Conversely, Th1-associated IFN-γ production was increased. An irreversible inhibitor of ITK, ibrutinib, blocked ex vivo Th17 generation and IL-17A production, conversely augmented FOXP3 expression only at low doses in Treg cultures. Finally, we analyzed peripheral ILC populations and observed a relative decrease in ILC2 and ILC3 frequency in our ITK-deficient patient., Conclusions: To our knowledge, this is the first report showing that both genetic and chemical inhibition of ITK result in reduced Th17 generation and function in humans. We also report, for the first time, a reduction in ILC2 and ILC3 populations in an ITK-deficient human patient.
- Published
- 2019
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19. Proteome Analysis of Human Neutrophil Granulocytes From Patients With Monogenic Disease Using Data-independent Acquisition.
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Grabowski P, Hesse S, Hollizeck S, Rohlfs M, Behrends U, Sherkat R, Tamary H, Ünal E, Somech R, Patıroğlu T, Canzar S, van der Werff Ten Bosch J, Klein C, and Rappsilber J
- Subjects
- Base Sequence, Humans, RNA, Messenger genetics, RNA, Messenger metabolism, Disease genetics, Neutrophils metabolism, Proteome metabolism, Proteomics
- Abstract
Neutrophil granulocytes are critical mediators of innate immunity and tissue regeneration. Rare diseases of neutrophil granulocytes may affect their differentiation and/or functions. However, there are very few validated diagnostic tests assessing the functions of neutrophil granulocytes in these diseases. Here, we set out to probe omics analysis as a novel diagnostic platform for patients with defective differentiation and function of neutrophil granulocytes. We analyzed highly purified neutrophil granulocytes from 68 healthy individuals and 16 patients with rare monogenic diseases. Cells were isolated from fresh venous blood (purity >99%) and used to create a spectral library covering almost 8000 proteins using strong cation exchange fractionation. Patient neutrophil samples were then analyzed by data-independent acquisition proteomics, quantifying 4154 proteins in each sample. Neutrophils with mutations in the neutrophil elastase gene ELANE showed large proteome changes that suggest these mutations may affect maturation of neutrophil granulocytes and initiate misfolded protein response and cellular stress mechanisms. In contrast, only few proteins changed in patients with leukocyte adhesion deficiency (LAD) and chronic granulomatous disease (CGD). Strikingly, neutrophil transcriptome analysis showed no correlation with its proteome. In case of two patients with undetermined genetic causes, proteome analysis guided the targeted genetic diagnostics and uncovered the underlying genomic mutations. Data-independent acquisition proteomics may help to define novel pathomechanisms in neutrophil diseases and provide a clinically useful diagnostic dimension., (© 2019 Grabowski et al.)
- Published
- 2019
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20. Alternative Splicing Rescues Loss of Common Gamma Chain Function and Results in IL-21R-like Deficiency.
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Illig D, Navratil M, Kelečić J, Conca R, Hojsak I, Jadrešin O, Ćorić M, Vuković J, Rohlfs M, Hollizeck S, Bohne J, Klein C, and Kotlarz D
- Subjects
- Alternative Splicing, B-Lymphocytes immunology, Child, Preschool, Cholangitis genetics, Cholangitis immunology, Croatia, Cryptosporidiosis genetics, Cryptosporidiosis immunology, Diarrhea genetics, Diarrhea immunology, Humans, Interleukin-21 Receptor alpha Subunit deficiency, Interleukin-21 Receptor alpha Subunit immunology, Male, Respiratory Tract Infections genetics, Respiratory Tract Infections immunology, Severe Combined Immunodeficiency immunology, Interleukin-21 Receptor alpha Subunit genetics, Severe Combined Immunodeficiency genetics
- Abstract
Inborn errors in interleukin 2 receptor, gamma (IL2RG) perturb signaling of the common gamma chain family cytokines and cause severe combined immunodeficiency (SCID). Here, we report two brothers suffering from chronic cryptosporidiosis, severe diarrhea, and cholangitis. Pan T, B, and NK cell numbers were normal, but immunophenotyping revealed defective B cell differentiation. Using whole exome sequencing, we identified a base pair deletion in the first exon of IL2RG predicted to cause a frameshift and premature stop. However, flow cytometry revealed normal surface expression of the IL-2Rγ chain. While IL-2, IL-7, and IL-15 signaling showed only mild defects of STAT5 phosphorylation in response to the respective cytokines, IL-4- and IL-21-induced phosphorylation of STAT3 and STAT6 was markedly reduced. Examination of RNA isoforms detected alternative splicing downstream of IL2RG exon 1 in both patients resulting in resolution of the predicted frameshift and 16 mutated amino acids. In silico modeling suggested that the IL-2Rγ mutation reduces the stabilization of IL-4 and IL-21 cytokine binding by affecting the N-terminal domain of the IL-2Rγ. Thus, our study shows that IL2RG deficiency can be associated with differential signaling defects. Confounding effects of alternative splicing may partially rescue genetic defects and should be considered in patients with inborn errors of immunity.
- Published
- 2019
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21. Human RIPK1 deficiency causes combined immunodeficiency and inflammatory bowel diseases.
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Li Y, Führer M, Bahrami E, Socha P, Klaudel-Dreszler M, Bouzidi A, Liu Y, Lehle AS, Magg T, Hollizeck S, Rohlfs M, Conca R, Field M, Warner N, Mordechai S, Shteyer E, Turner D, Boukari R, Belbouab R, Walz C, Gaidt MM, Hornung V, Baumann B, Pannicke U, Al Idrissi E, Ali Alghamdi H, Sepulveda FE, Gil M, de Saint Basile G, Hönig M, Koletzko S, Muise AM, Snapper SB, Schwarz K, Klein C, and Kotlarz D
- Subjects
- B-Lymphocytes immunology, B-Lymphocytes pathology, Epithelial Cells immunology, Epithelial Cells pathology, Female, HCT116 Cells, HEK293 Cells, Humans, Male, Mutation, NF-kappa B genetics, NF-kappa B immunology, T-Lymphocytes immunology, T-Lymphocytes pathology, Cell Differentiation genetics, Cell Differentiation immunology, Immunity, Mucosal genetics, Inflammatory Bowel Diseases genetics, Inflammatory Bowel Diseases immunology, Inflammatory Bowel Diseases pathology, Intestinal Mucosa immunology, Intestinal Mucosa pathology, Receptor-Interacting Protein Serine-Threonine Kinases deficiency, Receptor-Interacting Protein Serine-Threonine Kinases immunology, Severe Combined Immunodeficiency genetics, Severe Combined Immunodeficiency immunology, Severe Combined Immunodeficiency pathology
- Abstract
Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) is a critical regulator of cell death and inflammation, but its relevance for human disease pathogenesis remains elusive. Studies of monogenic disorders might provide critical insights into disease mechanisms and therapeutic targeting of RIPK1 for common diseases. Here, we report on eight patients from six unrelated pedigrees with biallelic loss-of-function mutations in RIPK1 presenting with primary immunodeficiency and/or intestinal inflammation. Mutations in RIPK1 were associated with reduced NF-κB activity, defective differentiation of T and B cells, increased inflammasome activity, and impaired response to TNFR1-mediated cell death in intestinal epithelial cells. The characterization of RIPK1-deficient patients highlights the essential role of RIPK1 in controlling human immune and intestinal homeostasis, and might have critical implications for therapies targeting RIPK1., Competing Interests: The authors declare no conflict of interest.
- Published
- 2019
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22. Intestinal Inflammation and Dysregulated Immunity in Patients With Inherited Caspase-8 Deficiency.
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Lehle AS, Farin HF, Marquardt B, Michels BE, Magg T, Li Y, Liu Y, Ghalandary M, Lammens K, Hollizeck S, Rohlfs M, Hauck F, Conca R, Walz C, Weiss B, Lev A, Simon AJ, Groß O, Gaidt MM, Hornung V, Clevers H, Yazbeck N, Hanna-Wakim R, Shouval DS, Warner N, Somech R, Muise AM, Snapper SB, Bufler P, Koletzko S, Klein C, and Kotlarz D
- Subjects
- Age of Onset, Animals, Apoptosis, Autoimmune Lymphoproliferative Syndrome therapy, Biopsy, Colitis, Ulcerative pathology, Colitis, Ulcerative therapy, Endoscopy, Gastrointestinal, Epithelial Cells pathology, Genetic Predisposition to Disease, Heredity, Humans, Inflammasomes immunology, Inflammation Mediators immunology, Intestines pathology, Lymphocytes pathology, Mice, Knockout, Mutation, Phenotype, Autoimmune Lymphoproliferative Syndrome genetics, Autoimmune Lymphoproliferative Syndrome immunology, Caspase 8 genetics, Colitis, Ulcerative genetics, Colitis, Ulcerative immunology, Epithelial Cells immunology, Intestines immunology, Lymphocytes immunology
- Published
- 2019
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23. Correction to: Novel Mutations in RASGRP1 Are Associated with Immunodeficiency, Immune Dysregulation, and EBV-Induced Lymphoma.
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Somekh I, Marquardt B, Liu Y, Rohlfs M, Hollizeck S, Karakukcu M, Unal E, Yilmaz E, Patiroglu T, Cansever M, Frizinsky S, Vishnvenskia-Dai V, Rechavi E, Stauber T, Simon AJ, Lev A, Klein C, Kotlarz D, and Somech R
- Abstract
The original version of this article unfortunately contained mistakes in Author's name, in Table 1 and in result section.
- Published
- 2018
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24. Novel Mutations in RASGRP1 are Associated with Immunodeficiency, Immune Dysregulation, and EBV-Induced Lymphoma.
- Author
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Somekh I, Marquardt B, Liu Y, Rohlfs M, Hollizeck S, Karakukcu M, Unal E, Yilmaz E, Patiroglu T, Cansever M, Frizinsky S, Vishnvenska-Dai V, Rechavi E, Stauber T, Simon AJ, Lev A, Klein C, Kotlarz D, and Somech R
- Subjects
- Alleles, Autoimmunity, Biomarkers, CRISPR-Cas Systems, Cell Differentiation genetics, Cell Differentiation immunology, Cell Line, Tumor, Child, Preschool, DNA Mutational Analysis, Female, Gene Expression, Gene Knockdown Techniques, Genotype, Humans, Immunologic Deficiency Syndromes metabolism, Infant, Infant, Newborn, Lymphoma metabolism, Lymphoproliferative Disorders etiology, Lymphoproliferative Disorders metabolism, Male, Pedigree, Exome Sequencing, DNA-Binding Proteins genetics, Disease Susceptibility, Epstein-Barr Virus Infections complications, Guanine Nucleotide Exchange Factors genetics, Immunologic Deficiency Syndromes etiology, Immunomodulation genetics, Lymphoma etiology, Mutation
- Abstract
Purpose: RAS guanyl-releasing protein 1 (RASGRP1) deficiency has recently been shown to cause a primary immunodeficiency (PID) characterized by CD4
+ T cell lymphopenia and Epstein-Barr virus (EBV)-associated B cell lymphoma. Our report of three novel patients widens the scope of RASGRP1 deficiency by providing new clinical and immunological insights on autoimmunity, immune cell development, and predisposition to lymphoproliferative disease., Methods: One patient of Turkish origin (P1) and two Palestinian patients (P2, P3) were evaluated for immunodeficiency. To decipher the molecular cause of disease, whole exome sequencing was conducted. Identified mutations were validated by immunological and biochemical assays., Results: We report three patients presenting with similar clinical characteristics of immunodeficiency and EBV-associated lymphoproliferative disease. In addition, P2 and P3 exhibited overt autoimmune manifestations. Genetic screening identified two novel loss-of-function mutations in RASGRP1. Immunoblotting and active Ras pull-down assays confirmed perturbed ERK1/2 signaling and reduced Ras-GTPase activity in heterologous Jurkat cells with ectopic expression of RASGRP1 mutants. All three patients had CD4+ T cell lymphopenia. P2 and P3 showed decreased mitogen-induced lymphocyte proliferation, reduced T cell receptor excision circles, abnormal T cell receptor (TCR) Vβ repertoires, and increased frequencies of TCRγδ cells. TCR gamma repertoire diversity was significantly reduced with a remarkable clonal expansion., Conclusions: RASGRP1 deficiency is associated with life-threatening immune dysregulation, severe autoimmune manifestations, and susceptibility to EBV-induced B cell malignancies. Early diagnosis is critical and hematopoietic stem cell transplantation might be considered as curative treatment.- Published
- 2018
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25. Human TGF-β1 deficiency causes severe inflammatory bowel disease and encephalopathy.
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Kotlarz D, Marquardt B, Barøy T, Lee WS, Konnikova L, Hollizeck S, Magg T, Lehle AS, Walz C, Borggraefe I, Hauck F, Bufler P, Conca R, Wall SM, Schumacher EM, Misceo D, Frengen E, Bentsen BS, Uhlig HH, Hopfner KP, Muise AM, Snapper SB, Strømme P, and Klein C
- Subjects
- DNA Mutational Analysis, Female, Humans, Inflammatory Bowel Diseases pathology, Male, Pedigree, Severity of Illness Index, Brain Diseases complications, Brain Diseases genetics, Inflammatory Bowel Diseases complications, Inflammatory Bowel Diseases genetics, Transforming Growth Factor beta1 genetics
- Abstract
Transforming growth factor (TGF)-β1 (encoded by TGFB1) is the prototypic member of the TGF-β family of 33 proteins that orchestrate embryogenesis, development and tissue homeostasis
1,2 . Following its discovery3 , enormous interest and numerous controversies have emerged about the role of TGF-β in coordinating the balance of pro- and anti-oncogenic properties4,5 , pro- and anti-inflammatory effects6 , or pro- and anti-fibrinogenic characteristics7 . Here we describe three individuals from two pedigrees with biallelic loss-of-function mutations in the TGFB1 gene who presented with severe infantile inflammatory bowel disease (IBD) and central nervous system (CNS) disease associated with epilepsy, brain atrophy and posterior leukoencephalopathy. The proteins encoded by the mutated TGFB1 alleles were characterized by impaired secretion, function or stability of the TGF-β1-LAP complex, which is suggestive of perturbed bioavailability of TGF-β1. Our study shows that TGF-β1 has a critical and nonredundant role in the development and homeostasis of intestinal immunity and the CNS in humans.- Published
- 2018
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26. Myb-like, SWIRM, and MPN domains 1 (MYSM1) deficiency: Genotoxic stress-associated bone marrow failure and developmental aberrations.
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Bahrami E, Witzel M, Racek T, Puchałka J, Hollizeck S, Greif-Kohistani N, Kotlarz D, Horny HP, Feederle R, Schmidt H, Sherkat R, Steinemann D, Göhring G, Schlegelbeger B, Albert MH, Al-Herz W, and Klein C
- Subjects
- Cells, Cultured, Consanguinity, DNA Repair genetics, DNA-Binding Proteins genetics, Genome-Wide Association Study, Genotype, Histones metabolism, Humans, Pedigree, Sequence Deletion genetics, Trans-Activators, Transcription Factors genetics, Ubiquitin-Specific Proteases, Ubiquitination, p38 Mitogen-Activated Protein Kinases metabolism, Bone Marrow Diseases immunology, DNA Damage immunology, DNA-Binding Proteins metabolism, Developmental Disabilities immunology, Hematopoiesis, Hematopoietic Stem Cell Transplantation, Immunologic Deficiency Syndromes immunology, Transcription Factors metabolism
- Abstract
Background: Myb-like, SWIRM, and MPN domains 1 (MYSM1) is a transcriptional regulator mediating histone deubiquitination. Its role in human immunity and hematopoiesis is poorly understood., Objectives: We sought to investigate the clinical, cellular, and molecular features in 2 siblings presenting with progressive bone marrow failure (BMF), immunodeficiency, and developmental aberrations., Methods: We performed genome-wide homozygosity mapping, whole-exome and Sanger sequencing, immunophenotyping studies, and analysis of genotoxic stress responses. p38 activation, reactive oxygen species levels, rate of apoptosis and clonogenic survival, and growth in immune and nonimmune cells were assessed. The outcome of allogeneic hematopoietic stem cell transplantation (HSCT) was monitored., Results: We report 2 patients with progressive BMF associated with myelodysplastic features, immunodeficiency affecting B cells and neutrophil granulocytes, and complex developmental aberrations, including mild skeletal anomalies, neurocognitive developmental delay, and cataracts. Whole-exome sequencing revealed a homozygous premature stop codon mutation in the gene encoding MYSM1. MYSM1-deficient cells are characterized by increased sensitivity to genotoxic stress associated with sustained induction of phosphorylated p38 protein, increased reactive oxygen species production, and decreased survival following UV light-induced DNA damage. Both patients were successfully treated with allogeneic HSCT with sustained reconstitution of hematopoietic defects., Conclusions: Here we show that MYSM1 deficiency is associated with developmental aberrations, progressive BMF with myelodysplastic features, and increased susceptibility to genotoxic stress. HSCT represents a curative therapy for patients with MYSM1 deficiency., (Copyright © 2017 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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