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Marco Dott. Ladetto

Background
MCL is a well-defined entity accounting for approximately 5% of non-Hodgkin lymphomas (NHL). Despite its poor prognosis the clinical outcome of MCL has improved over time both for progression free survival (PFS) and overall survival (OS).1 The most powerful prognostic tool in MCL is Real Time Quantitative (RQ)-PCR-based MRD2,3: such a tool provides a powerful independent predictor, which is currently investigated as a secondary endpoint in most ongoing MCL trials (e.g. IIL-MCL0208, EudraCT 2009-012807-25; FIL-R2B, EudraCT 2011-005461-21; FIL-RBAC500, NCT01662050; European MCL Network MCL-R2-Elderly, EudraCT 2012-002542-20). Some studies also offer a MRD-tailored treatment in specific therapeutic settings.4 PCR-based MRD is highly sensitive and reproducible, however it has some limitations, including marker identification failure and suboptimal sensitivity in some patients. High-throughput sequencing (HTS) might overcome these limitations, increase sensitivity and applicability and allow better characterization and monitoring of resistant clones that may contribute to disease progression and chemorefractoriness.5  

Study aims 
To develop an effective approach for MRD detection using HTS-based sequencing for the immunoglobulin heavy (IGH) and light chain genes. The approach will be similar to those already reported by Faham et al.5 and Ladetto et al.6 and will be pursued in the context of a large European multi-laboratory context (ESLHO Euro Clonality MRD NGS, http://www.euroclonality.org/). Critical aims will be to obtain: a) a tumor-specific marker in > 90% of MCL cases; b) a sensitivity of at least 1.0 EE-06. In parallel, a novel HTS-based approach will be developed for detection at diagnosis and subsequent monitoring of gene mutations targeting in MCL (most frequently mutated genes: ATM, CCND1, TP53, BIRC3, TLR2, WHSC1, MLL2, MEF2B, NOTCH1, NOTCH2).7 Using the clinical results of the IIL-MCL0208 study, the MRD results generated using classical RQ-PCR-based MRD and those obtained from the planned ancillary biological sub-studies, we aim at the following: a) comparing MRD results from IGH-based HTS with those of standard IGH-based RQ-PCR (already financed in the IIL-MCL0208 study), in terms of sensitivity, specificity and discordances between the two methods; b) correlating HTS-based MRD results with candidate biomarkers analyzed at diagnosis in the context of ancillary biological sub-studies, particularly flow cytometry analysis; c) clinically validating IGH-based HTS MRD results in terms of predictive value for PFS and OS, with the ultimate aim of substituting RQ-PCR-based MRD with the novel HTS approach in future studies; d) correlating IGH-based HTS MRD results with gene mutation HTS data, to dissect the sub-clonal nature of MCL. This last aim will be performed on diagnostic and follow-up samples to assess whether different treatment approaches might act as selective drivers of specific mutational patterns and whether the emergence of specific mutated sub-clones might be associated with the development of resistance to treatment.

Expected outcomes
1) to develop an effective approach to IGH-based HTS MRD detection that might result superior to RQ-PCR in terms of target identification, sensitivity and reproducibility;
2) to validate this approach in the context of a randomized clinical trial  (IIL-MCL0208) using RQ-PCR and flow cytometry as "gold standard"
3) to verify whether emergence/persistence of clones carrying specific adverse mutations might impact the  outcome of MCL patients 

  1. Dreyling M, Hiddemann W. Current treatment standards and emerging strategies in mantle cell lymphoma. Hematology Am Soc Hematol Educ Program 2009;542-551.
  2. Pott C, Hoster E, Delfau-Larue MH, Beldjord K, Böttcher S, Asnafi V, et al. Molecular remission is an independent predictor of clinical outcome in patients with mantle cell lymphoma after combined immunochemotherapy: a European MCL intergroup study. Blood. 2010 Apr 22;115(16):3215-23. doi: 10.1182/blood-2009-06-230250.
  3. Pott C, Hoster E, Beldjord K, Macintyre AE, Böttcher S, Asnafi V, et al. R-CHOP/R-DHAP Compared to R-CHOP Induction Followed by High Dose Therapy with Autologous Stem Cell Transplantation Induces Higher Rates of Molecular Remission In MCL: Results of the MCL Younger Intergroup Trial of the European MCL Network. Blood (ASH Annual Meeting Abstracts), Nov 2010; 116: 965.
  4. Andersen NS, Pedersen LB, Laurell A, Elonen E, Kolstad A, Boesen AM, et al. Pre-emptive treatment with rituximab of molecular relapse after autologous stem cell transplantation in mantle cell lymphoma. J Clin Oncol. 2009 Sep 10;27(26):4365-70. doi: 10.1200/JCO.2008.21.3116.
  5. Faham M, Zheng J, Moorhead M, Carlton VE, Stow P, Coustan-Smith E et al. Deep-sequencing approach for minimal residual disease detection in acute lymphoblastic leukemia. Blood 2012; 120: 5173-5180.
  6. Ladetto M, Brüggemann M, Monitillo L, Ferrero S, Pepin F, Drandi D, et al. Next-generation sequencing and real-time quantitative PCR for minimal residual disease detection in B-cell disorders. Leukemia. 2013 Dec 17. doi: 10.1038/leu.2013.375
  7. Beà S, Valdés-Mas R, Navarro A, Salaverria I, Martín-Garcia D, Jares P, et al. Landscape of somatic mutations and clonal evolution in mantle cell lymphoma. Proc Natl Acad Sci U S A. 2013 Nov 5;110(45):18250-5. doi: 10.1073/pnas.1314608110. 


Laboratory of Hematology, Molecular Biology

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Scientific Director:
Marco Ladetto, MD        marco.ladetto@unito.it

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Lab Team:

Daniela Barbero, PhD daniela.barbero@unito.it
Elisa Bernocco, MD      elisa.bernocco@libero.it
Daniela Drandi, PhD     daniela.drandi@unito.it
Simone Ferrero, MD simone.ferrero@unito.it
Elisa Genuardi, PhD elisa.genuardi@unito.it
Paola Ghione, MD paolaghione212@gmail.com
Barbara Mantoan, PhD barbara.mantoan@unito.it
Luigia Monitillo, PhD luigia.monitillo@unito.it

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Last update: 21/09/2018 23:32
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