Andrew Y Shuen1,2, Stella Lanni2, Gagan B Panigrahi2, Melissa Edwards2, Lisa Yu2, Brittany B Campbell1,2, Ariane Mandel2, Cindy Zhang2, Nataliya Zhukova1,2, Musa Alharbi3, Mark Bernstein4, Daniel C Bowers5,6, Sara Carroll7, Kristina A Cole8, Shlomi Constantini9,10, Bruce Crooks4, Rina Dvir10,11, Roula Farah12, Nobuko Hijiya13, Ben George14, Theodore W Laetsch5,6, Valerie Larouche15, Scott Lindhorst16, Rebecca C Luiten17, Vanan Magimairajan18, Gary Mason19, Warren Mason1,20, Oz Mordechai21, Naureen Mushtaq22, Garth Nicholas23, Michael Oren24, Laura Palma25, Luis Alberto Pedroza26,27, Jagadeesh Ramdas28, David Samuel29, Kami Wolfe Schneider30,31, Andrea Seeley28, Kara Semotiuk32, Ashraf Shamvil33, David Sumerauer34, Helen Toledano11, Patrick Tomboc35, Margaret Wierman31, An Van Damme36, Yi-Yen Lee37, Michal Zapotocky1,2,34, Eric Bouffet1,2, Carol Durno2,32, Melyssa Aronson32, Steve Gallinger32,38, William D Foulkes39, David Malkin1,2, Uri Tabori1,2, Christopher E Pearson1,2. 1. 1 University of Toronto, Toronto, Ontario, Canada. 2. 2 The Hospital for Sick Children, Toronto, Ontario, Canada. 3. 3 King Fahad Medical City, Riyadh, Saudi Arabia. 4. 4 Dalhousie University Faculty of Medicine, Halifax, Nova Scotia, Canada. 5. 5 University of Texas Southwestern Medical Center, Dallas, TX. 6. 6 Children's Health, Dallas, TX. 7. 7 Cleveland Clinic, Weston FL. 8. 8 Children's Hospital of Philadelphia and University of Pennsylvania, Philadelphia, PA. 9. 9 Tel Aviv Sourasky Medical Center, Tel Aviv, Israel. 10. 10 Tel Aviv University, Tel Aviv, Israel. 11. 11 Schneider Children's Medical Center of Israel, Petach Tikva, Israel. 12. 12 Saint George Hospital University Medical Center, Beirut, Lebanon. 13. 13 Ann & Robert H. Lurie Children's Hospital/Northwestern University Feinberg School of Medicine, Chicago, IL. 14. 14 Medical College of Wisconsin, Milwaukee, WI. 15. 15 Université Laval, Quebec City, Quebec, Canada. 16. 16 Medical University of South Carolina, Charleston, SC. 17. 17 Banner MD Anderson Cancer Center, Gilbert, AZ. 18. 18 University of Manitoba, Winnipeg, Manitoba, Canada. 19. 19 Children's Hospital of Pittsburgh of University of Pittsburgh Medical Center, Pittsburgh, PA. 20. 20 Princess Margaret Cancer Centre, Toronto, Ontario, Canada. 21. 21 Rambam Health Care Campus, Haifa, Israel. 22. 22 Aga Khan University Hospital, Karachi, Pakistan. 23. 23 Ottawa Hospital Research Institute, Ottawa, Ontario, Canada. 24. 24 Sheba Medical Center, Tel Hashomer, Israel. 25. 25 McGill University Health Centre, Montréal, Quebec, Canada. 26. 26 Baylor College of Medicine and Texas Children's Hospital, Houston, TX. 27. 27 Universidad San Francisco de Quito, Quito, Ecuador. 28. 28 Geisinger Medical Center, Danville, PA. 29. 29 Valley Children's Hospital, Madera, CA. 30. 30 Children's Hospital Colorado, Aurora, CO. 31. 31 University of Colorado, Anschutz Medical Campus, Aurora, CO. 32. 32 Mount Sinai Hospital, Toronto, Ontario, Canada. 33. 33 Children Cancer Hospital, Karachi, Pakistan. 34. 34 University Hospital Motol, Charles University, Prague, Czech Republic. 35. 35 Ruby Memorial Hospital, Morgantown, WV. 36. 36 Université Catholique de Louvain, Brussels, Belgium. 37. 37 Taipei Veterans General Hospital, Taipei, Republic of China. 38. 38 Toronto General Hospital, Toronto, Ontario, Canada. 39. 39 McGill University, Montréal, Quebec, Canada.
Abstract
PURPOSE: Constitutional mismatch repair deficiency (CMMRD) is a highly penetrant cancer predisposition syndrome caused by biallelic mutations in mismatch repair (MMR) genes. As several cancer syndromes are clinically similar, accurate diagnosis is critical to cancer screening and treatment. As genetic diagnosis is confounded by 15 or more pseudogenes and variants of uncertain significance, a robust diagnostic assay is urgently needed. We sought to determine whether an assay that directly measures MMR activity could accurately diagnose CMMRD. PATIENTS AND METHODS: In vitro MMR activity was quantified using a 3'-nicked G-T mismatched DNA substrate, which requires MSH2-MSH6 and MLH1-PMS2 for repair. We quantified MMR activity from 20 Epstein-Barr virus-transformed lymphoblastoid cell lines from patients with confirmed CMMRD. We also tested 20 lymphoblastoid cell lines from patients who were suspected for CMMRD. We also characterized MMR activity from patients with neurofibromatosis type 1, Li-Fraumeni syndrome, polymerase proofreading-associated cancer syndrome, and Lynch syndrome. RESULTS: All CMMRD cell lines had low MMR activity (n = 20; mean, 4.14 ± 1.56%) relative to controls (n = 6; mean, 44.00 ± 8.65%; P < .001). Repair was restored by complementation with the missing protein, which confirmed MMR deficiency. All cases of patients with suspected CMMRD were accurately diagnosed. Individuals with Lynch syndrome (n = 28), neurofibromatosis type 1 (n = 5), Li-Fraumeni syndrome (n = 5), and polymerase proofreading-associated cancer syndrome (n = 3) had MMR activity that was comparable to controls. To accelerate testing, we measured MMR activity directly from fresh lymphocytes, which yielded results in 8 days. CONCLUSION: On the basis of the current data set, the in vitro G-T repair assay was able to diagnose CMMRD with 100% specificity and sensitivity. Rapid diagnosis before surgery in non-neoplastic tissues could speed proper therapeutic management.
PURPOSE: Constitutional mismatch repair deficiency (CMMRD) is a highly penetrant cancer predisposition syndrome caused by biallelic mutations in mismatch repair (MMR) genes. As several cancer syndromes are clinically similar, accurate diagnosis is critical to cancer screening and treatment. As genetic diagnosis is confounded by 15 or more pseudogenes and variants of uncertain significance, a robust diagnostic assay is urgently needed. We sought to determine whether an assay that directly measures MMR activity could accurately diagnose CMMRD. PATIENTS AND METHODS: In vitro MMR activity was quantified using a 3'-nicked G-T mismatched DNA substrate, which requires MSH2-MSH6 and MLH1-PMS2 for repair. We quantified MMR activity from 20 Epstein-Barr virus-transformed lymphoblastoid cell lines from patients with confirmed CMMRD. We also tested 20 lymphoblastoid cell lines from patients who were suspected for CMMRD. We also characterized MMR activity from patients with neurofibromatosis type 1, Li-Fraumeni syndrome, polymerase proofreading-associated cancer syndrome, and Lynch syndrome. RESULTS: All CMMRD cell lines had low MMR activity (n = 20; mean, 4.14 ± 1.56%) relative to controls (n = 6; mean, 44.00 ± 8.65%; P < .001). Repair was restored by complementation with the missing protein, which confirmed MMR deficiency. All cases of patients with suspected CMMRD were accurately diagnosed. Individuals with Lynch syndrome (n = 28), neurofibromatosis type 1 (n = 5), Li-Fraumeni syndrome (n = 5), and polymerase proofreading-associated cancer syndrome (n = 3) had MMR activity that was comparable to controls. To accelerate testing, we measured MMR activity directly from fresh lymphocytes, which yielded results in 8 days. CONCLUSION: On the basis of the current data set, the in vitro G-T repair assay was able to diagnose CMMRD with 100% specificity and sensitivity. Rapid diagnosis before surgery in non-neoplastic tissues could speed proper therapeutic management.
Authors: Jiil Chung; Yosef E Maruvka; Gad Getz; Uri Tabori; Sumedha Sudhaman; Jacalyn Kelly; Nicholas J Haradhvala; Vanessa Bianchi; Melissa Edwards; Victoria J Forster; Nuno M Nunes; Melissa A Galati; Martin Komosa; Shriya Deshmukh; Vanja Cabric; Scott Davidson; Matthew Zatzman; Nicholas Light; Reid Hayes; Ledia Brunga; Nathaniel D Anderson; Ben Ho; Karl P Hodel; Robert Siddaway; A Sorana Morrissy; Daniel C Bowers; Valérie Larouche; Annika Bronsema; Michael Osborn; Kristina A Cole; Enrico Opocher; Gary Mason; Gregory A Thomas; Ben George; David S Ziegler; Scott Lindhorst; Magimairajan Vanan; Michal Yalon-Oren; Alyssa T Reddy; Maura Massimino; Patrick Tomboc; An Van Damme; Alexander Lossos; Carol Durno; Melyssa Aronson; Daniel A Morgenstern; Eric Bouffet; Annie Huang; Michael D Taylor; Anita Villani; David Malkin; Cynthia E Hawkins; Zachary F Pursell; Adam Shlien; Thomas A Kunkel Journal: Cancer Discov Date: 2020-12-18 Impact factor: 39.397
Authors: Richard Gallon; Barbara Mühlegger; Sören-Sebastian Wenzel; Harsh Sheth; Christine Hayes; Stefan Aretz; Karin Dahan; William Foulkes; Christian P Kratz; Tim Ripperger; Amedeo A Azizi; Hagit Baris Feldman; Anne-Laure Chong; Ugur Demirsoy; Benoît Florkin; Thomas Imschweiler; Danuta Januszkiewicz-Lewandowska; Stephan Lobitz; Michaela Nathrath; Hans-Jürgen Pander; Vanesa Perez-Alonso; Claudia Perne; Iman Ragab; Thorsten Rosenbaum; Daniel Rueda; Markus G Seidel; Manon Suerink; Julia Taeubner; Stefanie-Yvonne Zimmermann; Johannes Zschocke; Gillian M Borthwick; John Burn; Michael S Jackson; Mauro Santibanez-Koref; Katharina Wimmer Journal: Hum Mutat Date: 2019-03-06 Impact factor: 4.878
Authors: M Suerink; K Wimmer; L Brugieres; C Colas; R Gallon; T Ripperger; P R Benusiglio; E M A Bleiker; Z Ghorbanoghli; Y Goldberg; J C H Hardwick; M Kloor; M le Mentec; M Muleris; M Pineda; C Ruiz-Ponte; H F A Vasen Journal: Fam Cancer Date: 2020-07-02 Impact factor: 2.375
Authors: Maribel González-Acosta; Fátima Marín; Benjamin Puliafito; Gabriel Capellá; Marta Pineda; Nuria Bonifaci; Anna Fernández; Matilde Navarro; Hector Salvador; Francesc Balaguer; Silvia Iglesias; Angela Velasco; Elia Grau Garces; Victor Moreno; Luis Ignacio Gonzalez-Granado; Pilar Guerra-García; Rosa Ayala; Benoît Florkin; Christian Kratz; Tim Ripperger; Thorsten Rosenbaum; Danuta Januszkiewicz-Lewandowska; Amedeo A Azizi; Iman Ragab; Michaela Nathrath; Hans-Jürgen Pander; Stephan Lobitz; Manon Suerink; Karin Dahan; Thomas Imschweiler; Ugur Demirsoy; Joan Brunet; Conxi Lázaro; Daniel Rueda; Katharina Wimmer Journal: J Med Genet Date: 2019-09-07 Impact factor: 6.318