Projects

CENTRAL NERVOUS SYSTEM CLL, A RARE DISEASE WITH UNCLEAR MANAGEMENT

This is a retrospective data collection on CLL patients who developed CNS disease involvement.

CD-19-CAR-T CELLS IN PATIENTS WITH RICHTER’S TRANSFORMATION (RT)

Richter’s transformation (RT) is a very aggressive lymphoma with limited treatment options. The response rate and durability of CAR-T cells in RTs is unknown. Thus, we suggest a retrospective multicenter study to identify response to CAR-T cells in RT and define proportion of patients that are referred to allogeneic transplant following CAR-T.

CLONAL RELATIONSHIP IN RICHTER TRANSFORMATION (RT)

Riccardo Moia (Novara, Italy) launched an ERIC project aiming at studying biological and clinical features of RT stratified on their clonal relationship with the preceding CLL.

CARDIAC FUNCTION ASSESSMENT IN PATIENTS WITH CLL

ERIC is conducting a survey on if and how cardiac function is assessed in patients with CLL over disease course, in particular to understand how severe cardiac impairment is affecting treatment choice at the time of progression. We would be very grateful if you would complete this very brief survey to help ERIC gather information on cardiac function assessment, its role in patient management and the potential application in the design of interventional clinical trials with targeted agents.

CLL Patient Empowerment Program

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)     

Aliki Xochelli, Christina Karamanidou, Kostas Stamatopoulos (Affiliation: Centre for Research and Technology Hellas (CERTH) / Institute Of Applied Biosciences (INAB), Thessaloniki, (Greece)

Background      

A paradigm shift described in healthcare indicates that preventive, pre-emptive and predictive healthcare decisions should be made in a pervasive, participatory and personalized manner. It is widely accepted that such a paradigm shift is not possible using traditional tools and processes and requires the patient’s active engagement in the management of his/her own health. Evidently, patient empowerment towards self-management requires novel methodological and technological frameworks, well-designed educational programs that will familiarize patients, caregivers and healthcare professionals alike with the concept of co-decision, its scope and challenges but also its misuse. We herein envision patient empowerment as a four-arm, iterative process that involves patients, caregivers and healthcare professionals, namely physicians and nurses, and propose a framework for chronic lymphocytic leukemia (CLL), the most frequent adult hematologic cancer. Our CLL patient empowerment program aims to engage physicians and nurses, empower and support patients and caregivers and promote co-decisions between physicians and patients by establishing a common language and structured ways of communication.

Main objectives             

1. To explore CLL patients’ experience of illness (physical, emotional, social dimension), and specifically their understanding of the disease (physical dimension), their ways of coping, perceived impact etc. Also, their experience regarding treatment, their understanding and concerns as well as their relationship with physicians etc.
2. To explore health care professionals experience in treating patients with CLL and identify their needs and concerns with regards to communication within the context of patient-physician relationship.
3. To systematically record patients’ beliefs, knowledge and preferences regarding their illness and treatment, their mood, their satisfaction concerning medical information, their adherence to medication etc.
4. To systematically record patient reported outcomes regarding symptoms, adverse effects and life style changes throughout the disease course.
5. To develop an educational framework for (i) the enhancement of physicians communication skills and (ii) the enhancement of patients self assessment and self management and also their caregivers knowledge on CLL

Main activities               

• Face to face interviews/ focus groups with participants (health care professionals and patients/caregivers)
• Self-completed Questionnaires −
• Development of a CLL knowledge questionnaire and a treatment preference questionnaire for patients –
• Design, development and evaluation of tools that will support physician communication and patient empowerment

Project Start date           1 January 2017

Project Finish date         1 January 2020

Different phases within the project     

• Phase I- Needs assessment phase
• Phase II- Design and development of tools
• Phase III- Evaluation of tools

Name of sponsor (s) if any:  Janssen Global ; Gilead Sciences Hellas; Janssen Cilag Greece ;

Contact person (s) for further details:  Aliki Xochelli; aliki.xochelli@gmail.com; Christina Karamanidou; ckaramanidou@gmail.com

International CLL registry for spontaneous regression in CLL

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Ilaria Del Giudice, Emili Montserrat IDG: Affiliations; Division of Hematology, Department of Cellular Biotechnologies and Hematology, “Sapienza” University, Rome (Italy); Hospital Clinic Barcelona, Barcelona (Spain)

Background

Spontaneous regression in CLL in the absence of any previous treatment is exceptional and the biologic bases are unknown (Del Giudice et al. Blood 2009). Since the incidence of CLL spontaneous regression has been estimated about 1%, no single center will have many patients with this infrequent event. The proposal of an international registry for spontaneous regression in CLL has been launched among the ERIC members. The aim is to pool clinical data and biological information of these cases and making them accessible to many researchers for future studies.

Main objectives             

• Identify key differences with CLL cases which follow the usual natural history of the disease
• Formulate hypothesis on the pathogenesis of this phenomenon and on mechanism of control of CLL progression to more advanced stages.

Main activities

A dedicated form to collect clinical data (medical history, drug intake, biologic markers, etc.) is available in the website. Due to the rarity of this condition, the project is still open and running. So far, 24 new cases with spontaneous regression have been included in the registry, 15 from Spain, 7 from Italy and 2 from Sweden. A new call has been launched in these days to researchers interested in CLL.

In the case peripheral blood samples can be obtained, biological studies on the residual CLL clone and on the T cell populations will be performed.

Project Start date           15 February 2011

Project Finish date         15 December 2018

Different phases within the project

Phase I (collection of clinical data in the registry). Likely, the registry will be closed by the end of 2018.

Phase II (collection of peripheral blood samples for biologic studies) will be eventually opened afterwards.

Contact person (s) for further details: Ilaria Del Giudice, delgiudice@bce.uniroma1.it   

Obinutuzumab + Chlorambucil as front-line treatment in CLL

Observational study to assess the efficacy and safety of bendamustine plus rituximab in patients affected by chronic lymphocytic leukemia

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)     

Antonio Cuneo; Affiliation: University of Ferrara, Ferrara (Italy)

Background      

The efficacy and safety of the BR regimen in first line and in second line in the clinical practice is not defined

Main objectives             

The primary objective of the study is to observe the progression-free survival in CLL patients treated with first and second-line BR regimen.

Main activities  compilation of the CRF

Project Start date           10 January 2016

Project Finish date         Q1 2018

Different phases within the project     

1) Collection of the data

2) Writing of the manuscript

Name of sponsor (s) if any:    GIMEMA-ERIC

Contact person (s) for further details:  Antonio Cuneo; cut@unife.it

Prognostic and predictive value of complex karyotyping using chromosome banding analysis or microarray-based profiling in CLL

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Arnon Kater; Kostas Stamatopoulos; Affiliations: Academic Medical Center Amsterdam – University of Amsterdam, Amsterdam, (The Netherlands); Centre for Research and Technology Hellas (CERTH) / Institute Of Applied Biosciences (INAB), Thessaloniki, (Greece)

Background

Today, the ‘gold standard’ for genetic risk stratification in CLL is FISH analysis with probes for chromosomes 11q, 12, 13q and 17p complemented by mutational screening for the TP53 gene found to be recurrently mutated in CLL. Such screening (particularly for del17p and TP53 mutations) has helped to select patients with low versus high response chances to standard chemo-immunotherapy. This is especially relevant considering the rapid expansion of effective but highly expensive alternative treatment regimens in high-risk patients. Therefore, improved risk-stratification has become a priority in CLL patient management.

Studies from the early 1990s provided the first evidence for the turbulent genetic landscape of CLL as revealed by classic chromosome banding analysis (CBA). Against this evidence, FISH superseded CBA in the routine diagnostic setting as the latter was more laborious, considering the need of in vitro stimulation in order to obtain mitoses. That notwithstanding, FISH is assessing only selected chromosomal regions whereas CBA offers the opportunity to globally assess the karyotype of the malignant clone. Indeed, by providing a more comprehensive view of the genetic landscape, CBA may offer valuable complementary information to the routinely performed FISH analysis in CLL, potentially, improving risk stratification. This is especially so today, when effective and reliable cell stimulation protocols are available, overcoming the limitations of older protocols.

More recently, CBA in CLL has attracted great interest in view of reports indicating: complex karyotype (CK) as a potential predictive marker for refractoriness to ibrutinib, independently of the presence of the chromosome 17p deletion [del(17p)]; and, lack of any adverse prognostic effect of CK on either PFS or OS, irrespective of the presence of del(17p)/TP53mut, in idelalisib-treated patients. That said, the available evidence does not yet allow reaching definitive conclusions regarding either the precise role of signaling inhibitors at large in the management of CLL patients with CK or the relative value of one inhibitor over the other in this particular group of patients.

Altogether, these findings allude to the important potential role of comprehensive karyotype analysis for refined risk stratification in CLL. However, many challenges towards routine clinical application of CBA still need to be overcome, thus indicating the urgent need for rigorous definitions, particularly regarding which cytogenetic profiles exactly correspond to CK, validated methodologies and systematic investigation in large series.

Array-based analysis is another powerful technique capable of globally assessing the CLL genome. From a technical perspective, in contrast to CBA, no cell stimulation step is needed for array-based analysis which also offers increased detection depth regarding deletions or duplications. However, balanced aberrations cannot be detected by micro-arrays, which is not trivial, especially considering reports highlighting their potential prognostic impact on clinical outcome.

The application of array-based analysis in CLL not only revealed the well-known recurrent chromosome abnormalities but also recovered additional (recurrent) genetic alterations, including complex genomic profiles with 3 or more acquired copy number aberrations (aCNAs). Elevated CLL genomic complexity demonstrated by microarrays identified patients with aggressive CLL and short survival.

Main objectives             

1. To compare with regards to the detection rate of CK:

• different CBA stimulation protocols [CpG-oligodeoxynucleotides/interleukin 2 (CpG/IL2), phorbol-12-myristate-13-acetate (TPA) or both]
• CBA versus microarray analysis in well annotated patient samples.

2. To assess the prognostic and predictive relevance of various numerical cut-offs for cytogenetic aberrations (3 vs 4 vs 5 or more) for defining cytogenetic complexity and investigate the importance of the type of aberrations (i.e. structural versus numerical).
3. To assess the potential associations between CK and other genetic features of the malignant clones, particularly the presence of TP53 aberrations (deletion of chromosome 17p and/or TP53 mutation).
4. To determine the precise prognostic role of CK in risk stratification of patients with CLL.

Main Activities

• Gather and analyze CBA data as performed in labs through Europe
• Gather and analyze array data as performed in labs through Europe
• Define exact definitions of CK in CLL
• Compare the reproducibility of the 2 techniques

Project Start date           1 February 2016

Project Finish date         1 December 2018

Different phases within the project

• Analysis of CBA data from different labs in Europe
• Analysis of array data from different labs in Europe
• Harmonization activities
• CBA and array analysis of CLL patients treated with novel biological agents

Name of sponsor (s) if any

Janssen

Contact person (s) for further details

Arnon Kater; a.p.kater@amc.nl; Kostas Stamatopoulos; kostas.stamatopoulos@certh.gr

TP53 Variants Database in CLL 

Organizing centers
Brno: Jitka Malcikova, Sarka Pavlova, Sarka Pospisilova
Thessaloniki: Kostas Stamatopoulos, Anastasia Hadzidimitriou, Evangelia Minga
Collaborators:
Prof. Thierry Soussi (Uppsala, Paris)

Background
CLL was the first malignancy for which testing for the presence of TP53 defects to guide treatment decision-making was fully implemented in the clinics and the recommendations for TP53 analysis formulated by ERIC have been valid since 2012 (Pospisilova et al., Leukemia 2012; Malcikova et al., Leukemia 2018). This creates a unique opportunity to collect the list of TP53 variants obtained during routine TP53 screening and analyze the variant profile.

Main objectives
To create a publicly accessible CLL-specific database of TP53 variants.

Main activities

• Collecting the detailed information on TP53 variants in CLL patients from participating centers
• Curation of the data for non-pathogenic variants and/or potential SNPs
• Analyzing a profile of TP53 variants detected in CLL (update of Zenz et al., Leukemia 2010)
• Releasing the TP53 database
• Gathering clinical data either by connecting with the data submitted to ERIC CLL database or by providing clinical data independently
• Analyzing different TP53 profiles in the context of different clinical and biological variables (e.g. FISH, IGHV, treatment status, outcome…)

Addressed centers      All ERIC members, TP53 certification required

Project Start date        Spring 2021

Project Finish date     Not defined (ongoing database administration)

Different phases within the project:

• Addressing ERIC members to ascertain interest in participating in the project
• TP53 data collection
• TP53 data curation and functional anotation
• Upload to the ERIC CLL database and TP53 website and/or IARC database
• Data analysis
• Writing a manuscript

ERIC CLL Database

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)
Anastasia Chatzidimitriou; Carol Moreno; Affiliations: Institute of Applied Biosciences, Thessaloniki, Greece; Hospital Sant Pau, Barcelona, Spain

Background
Chronic Lymphocytic Leukemia (CLL) is a chronic malignancy of B-lymphocytes with a distinctive immunophenotype. The incidence of this disease is around 5 cases per 100,000 people per year. CLL prognosis is variable, and the disease is mostly incurable.

The availability of clinical and biological data of patients with CLL could facilitate CLL biological, translational, and clinical research across Europe, likely collected in a standardized fashion though flexible enough to be integrated with local existing registries and adapted to  project-specific data collection forms.  The European Research Initiative on CLL (ERIC) is an organization aimed at promoting research in CLL across Europe. ERIC is composed by more almost 1000 members belonging to several institutions within Europe as well as in all other continents. Hence, it is in a unique position to coordinate the collection, curation, storage and analysis of real-world evidence from patients with CLL as described above.

Main objectives

Primary

• To collect prospective and retrospective clinical and biological data of patients with CLL at the time of diagnosis and follow-up on a project basis

Secondary

• To develop a data management system and a database for handling and storage of clinicobiological data
• To provide longitudinal data for outcome studies
• To generate hypotheses regarding the association of potential prognostic variables with outcomes in this patient population
• To provide benchmark data about the impact of CLL on European health-systems

Main activities

Primary

• To collect basic information about patients with CLL (e.g. age, gender, comorbidity, performance status)
• To collect disease-related information (date of diagnosis, stage, biological parameters, complete blood cell count)
• To record data about CLL-directed treatment (e.g. types and routes of therapy), response, complications and adverse events
• To record relevant outcome data including survival (overall and progression-free, date of last follow up, death, cause of death)

Secondary

• To generate hypotheses regarding important disease characteristics, laboratory studies and therapies
• To define relevant parameters influencing CLL impact on health systems

Data collection will be conducted as either a retrospective or prospective medical record review as follows:

– All the registries and updates will be stored in a local repository in every participating center. In case, there is no registry currently available at a participating centre, a local registry will be set up. Informatics support will be provided to transfer the local registries into the novel local repository.

– A web-based interface provided by ERIC will be used for data registration and connected to the local repository. All records, and their regular updates, will be anonymized locally, uploaded and saved to the CLL-db periodically. It is important to note that the correspondence between original and anonymized records (as well as the ability to track changes) will be stored only at the local level. As such, the central CLL-db will not have the ability to track changes nor have access to the mapping knowledge.

– Queries to the central CLL-db will be allowed based on the project and will retrieve only aggregated, anonymized data.

– This system will require installation of the software locally and informatics support will be provided by ERIC  collaborating with the system administrator of each center.

Project Start date                  1 January 2016

Project Finish date                Not defined, on a project basis

Different phases within the project

• Local system requirements
• Data management system adaptation to local system requirements
• Data management system installation to the participating centres
• Data collection
• Data analysis

Contact person (s) for further details:  Anastasia Chatzidimitriou; achatzidimitriou@certh.gr

BcR IG sequence collection

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)     

Kostas Stamatopoulos; Andreas Agathangelidis; Affiliation: Centre for Research and Technology Hellas (CERTH) / Institute Of Applied Biosciences (INAB),  Thessaloniki , Greece

Background      

Immunogenetic analysis of the B cell receptor immunoglobulin (BcR IG) has been instrumental for understanding the pathogenesis of chronic lymphocytic leukemia (CLL) and has also helped in the classification of patients into groups with distinct prognosis. A highly restricted BcR IG gene repertoire was the first evidence for antigen selection in the natural history of CLL. Additional evidence was provided by the differential prognosis of cases with distinct somatic hypermutation load within the clonotypic immunoglobulin heavy variable (IGHV) genes. Yet, perhaps the strongest evidence for the importance of microenvironmental interactions in driving CLL development and evolution is the existence of subsets of patients with stereotyped BCRs, collectively accounting for a remarkable one-third of the entire cohort. These observations have been instrumental in shaping the notion that CLL ontogeny is functionally driven and dynamic, rather than a simple stochastic process.

Main objectives             

In collaboration with IMGT®, the International Immunogenetics Information System, this ERIC project entails the expansion and maintenance of the IMGT/CLL-DB (http://www.imgt.org/CLLDBInterface/query), a database holding primary IG sequences from patients with CLL to foster high-quality collaborative research on CLL and related disorders, particularly regarding the pathogenesis and molecular epidemiology of CLL.

Main activities

Main activities include: (i) data acquisition, (ii) data processing and curation, (iii) data upload on IMGT/CLL-DB, (iv) data analysis.

Project Start date           1 January 2018

Project Finish date         31 December 2025 

Different phases within the project     

Phase 1: data collection,

Phase 2: data curation,

Phase 3: upload to the IMGT/CLL-DB,

Phase 4: BcR IG repertoire analysis.

Name of sponsor (s) if any        

Janssen

Contact person (s) for further details  

Andreas Agathangelidis; aagathangelidis@gmail.com

Comparative analysis of target enrichment techniques for the detection of gene mutations in CLL

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Lesley Ann Sutton; Richard Rosenquist; Affiliations: Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden; Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden

Background

Targeted NGS, in which a select number of genes are sequenced, is emerging as a much more efficient means to identify genetic variants and contains many attractive features, including: 1) the capacity to custom design a gene panel and screen a large number of genes and samples simultaneously; 2) the ability to detect mutations present at low variant allelic frequencies due to the high sequence depth achievable; and 3) the possibility to analyse all coding exons within a gene, irrespective of its length. As a consequence, this methodology is now being considered within everyday clinical practice. However, with several enrichment techniques now available for targeted sequencing, prior to implementing such a workflow, questions that commonly arise concern the specific technique that should be chosen, whether differences in sensitivity/specificity exist between the various techniques, which genes should be analysed, the optimal number of samples to multiplex, the appropriate bioinformatics workflow etc.

Main objectives

Motivated by this unmet need and uncertainty, ERIC has launched an initiative aimed at simplifying the introduction of targeted NGS into clinical laboratories by identifying the pros or cons (if any) of various enrichment techniques and determining the most suitable for the detection of recurrent mutations in CLL as well as ensuring end-to-end harmonization in the diagnostic approach.

Main activities

Three enrichment techniques (Haloplex, Agilent; TruSeq, illumina; CLL MASTR plus, Multiplicom) will be compared. Each technique will be analyzed by at least 2 centers and all sequencing will be performed on the Illumina MiSeq instrument. Eleven genes recurrently mutated in CLL will be included in the analysis. Forty-eight CLL samples with matched germline DNA will be included in the study (all centers will analyse the same samples). To be included in the study a sample must be well characterized and carry a mutation in at least one of the selected genes.

Project Start date           1 May 2015

Project Finish date         30 November 2017

Different phases within the project

• Phase 1: Comparative analysis of target enrichment techniques;
• Phase 2: Large-scale validation of a gene panel for CLL diagnostics;
• Phase 3: Longitudinal studies using NGS technologies

Contact person (s) for further details:  Lesley Ann Sutton; lesley-ann.sutton@ki.se

MULTICENTER STUDY ON PROGNOSTIC AND PREDICTIVE IMPACT OF TP53 VARIANTS BELOW 10% VAF

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Sarka Pospisilova, Sarka Pavlova, Jitka Malcikova; Central European Institute of Technology (CEITEC), Masaryk University Brno; Department of Internal Medicine – Hematology and Oncology, University Hospital Brno; Czech Republic

Co-investigators

Stockholm, Sweden, Richard Rosenquist Brandell and Lesley-Ann Sutton; Copenhagen, Denmark, Carsten Utoft Niemann; Bellinzona, Switzerland, Davide Rossi; Milan, Italy, Silvia Bonfiglio and Paolo Ghia

Background

TP53 mutations in chronic lymphocytic leukemia (CLL) patients have independent negative impact on patients' prognosis and response to chemo-immunotherapy regimens. Patients with TP53 mutations unequivocally benefit from targeted therapy with novel agents, e.g. BCR or bcl-2 inhibitors. Moreover, cells with TP53 mutations often outcompete prevailing TP53-wild-type (TP53- wt) subclones under chemotherapy pressure which may lead to the expansion of minor TP53- mutated (TP53-mut) subclone(s) in disease relapse accompanied by rapid deterioration of the prognosis.
Currently ERIC recommends reporting of TP53 variants present in ≥10% variant allelic frequency (VAF). However, there is a strong need for future re-evaluation of this threshold, aiming to decrease risks for patients with smaller TP53-mut subclones.
A well-defined study collecting data from multiple centers may overcome drawbacks of single-center studies, especially low sample numbers in the studied subgroups, and provide robust conclusions. To reach this, NGS data from consecutive samples of CLL patients entering first-line
therapy will be collected from participating centers. One of the barriers both for performing multicenter study on clinical impact of minor TP53 variants and decreasing the reporting limit has been so far the lack of confidence and/or reliable next generation sequencing (NGS) methodology to detect variants below 10%. To avoid methodical bias, a methodical harmonization phase will be performed as a first step to ensure that all methodologies included in the study are equally able to detect minor mutations in reference samples.

Main Objectives

– To assess prognostic and predictive impact of low-VAF TP53 variants in patients entering first-line treatment

– To re-evaluate the cut-off for reporting of TP53 variants in CLL and, if needed, to update recommendations on minor TP53 variant detection, validation and reporting

– To compare results among laboratories performing NGS analysis of TP53 mutations in CLL with detection limit ≤1% VAF

Main Activities

– Inter-laboratory comparison of NGS results obtained from the set of reference samples

Addressed laboratories: Centers certified for TP53 analysis by ERIC

– Data collection – results of TP53 analysis using NGS with detection limit ≤1% VAF

Required data: Results of TP53 analysis using NGS methodology with detection limit ≤1% VAF.
Corresponding clinical and laboratory data (disease course and routine prognostic markers)

Sample set

Inclusion criteria

1) Consecutive set of samples from CLL patients entering first-line therapy.Minimal follow-up of 4 years (i.e. sampled before 2015).

2) Clinical and laboratory data available.

Exclusion criteria

1) The laboratory is not able to perform the NGS analysis reliably within given detection limit and avoid significant proportion of false negatives/positives.

2) Non-consecutive set of samples intentionally enriched for a specific group of patients, e.g. for retrospectively analyzed patients with later clonal expansion of TP53 defect, poor therapy response etc. This would introduce undesirable bias.

Different phases within the project

– Methodical harmonization (2019)
Sending reference samples with TP53 variants ≤10% – Q2 2019
Inter-laboratory comparison of NGS results obtained from the set of reference samples – Q4/2019

– Patient data collection and analysis (2020)
Collecting data for the study
Analysis of the data

Project Start date:    November 2018
Project Finish date: December 2020

Contact person (s) for further details: Sarka Pavlova; pavlova.sarka@fnbrno.cz

Prevalence of BTK and PLCϒ2 Mutations in Patients relapsing under Ibrutinib

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Paolo Ghia; IRCCS San Raffaele Scientific Institute, Milan, Italy.

Background

The first-in-class irreversible BTK inhibitor, ibrutinib, showed impressive efficacy in CLL leading to FDA and EMA approval. Late relapses on ibrutinib often associate with mutations within the Bruton’s tyrosine kinase (BTK) gene at the binding site of ibrutinib or within the PLCγ2 gene, that is located immediately downstream of BTK. Deep sequencing analysis confirmed that up to 80-85% of resistant cases harbor mutations within BTK and/or PLCG2 in a variable proportion of CLL cells in resistant cases, ranging from 0.1 to 80% [according to the estimated variant allele frequency (VAF)]. As CLL patients relapsing on ibrutinib represent a new unmet clinical need, thorough understanding of the mechanisms behind the development of resistance as well as the actual prevalence of the mechanisms so far identified is key to assist treatment decisions and improve the outcomes of such patients. To this aim, we propose this retrospective case-based study, aimed at characterizing the prevalence of BTK and PLCG2 mutations in CLL patients relapsing on ibrutinib and at investigating additional potential mechanisms of resistance.

Main objectives             

Primary Objective: To define the prevalence of BTK and/or PLCG2 mutations in chronic lymphocytic leukemia (CLL) patients relapsing under ibrutinib.

Secondary Objectives: To define the prevalence of BTK and/or PLCG2 mutations in CLL patients responding to ibrutinib after at least 1 year of treatment; To define the estimated variant allele frequency (VAF) in CLL patients relapsing under ibrutinib and in those responding to ibrutinib after at least 1 year of treatment.

Main activities

Targeted next-generation sequencing (NGS) of the BTK and PLCG2 genes.

Exploration of statistical associations with various clinicobiological features, including: demographic data; disease characteristics; previous treatment and ibrutinib treatment information.

Project Start date           November 2017

Project Finish date        June 2018

Different phases within the project     

Sample collection will start in November 2017. Library preparation and Sequencing will start in December 2017. Data analysis will start February 2018. Completion of the project is expected by June 2018.

Contact person (s) for further details:  Lydia Scarfò; scarfo.lydia@hsr.it; Silvia Bonfiglio; bonfiglio.silvia@hsr.it

Recurrent gene mutations in CLL: ERIC project in HARMONY

Project Leader (s), Coordinator (s) or/and Principal Investigator (s)

Lesley Ann Sutton; Richard Rosenquist; Affiliations: Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden; Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden

Background

Next-generation sequencing (NGS) technologies have proved invaluable for deciphering the molecular heterogeneity of chronic lymphocytic leukemia (CLL). Such studies have revealed a diverse pattern of mutations, with a limited number of frequently mutated genes identified followed by a seemingly endless tail of genes harboring mutations at an extremely low frequency. Evidence suggests that this observed biological heterogeneity is intricately linked to the heterogeneous pathological characteristics and clinical course evidenced in CLL patients. Although certain gene mutations have prognostic or even predictive capabilities, most gene mutations require additional rigorous validation in large, patient series, to not only discern their potential contribution (if any) to the pathobiology of CLL but also ascertain whether a particular gene mutation could aid in clinical decision-making, including therapy selection and response prediction.

Main objectives             

We will evaluate the mutational status of genes in a well-characterized cohort encompassing thousands (≈4000) of CLL patients through a coordinated, multi-center collaboration conducted under the auspices of the European Research Initiative in CLL (ERIC). More specifically, we will investigate: (i) the most recurrently mutated genes in CLL, namely NOTCH1, SF3B1, ATM, BIRC3, MYD88 and TP53; mutations within the TP53 and ATM genes have long been known to confer poor prognosis, however recurrent mutations within NOTCH1, SF3B1 and BIRC3 have more recently been reported to be associated with poor clinical outcome with higher frequencies in relapsing/refractory CLL and in Richter’s syndrome; and, (ii) an additional four genes, i.e. EGR2, NFKBIE, POT1 and XPO1; recurrent mutations in these genes have been associated with a dismal clinical course, although they have been less studied.

Main activities

1. Evaluation of the mutational status of NOTCH1, SF3B1, ATM, BIRC3, MYD88, TP53, EGR2, NFKBIE, POT1 and XPO1 in a large series of CLL cases, well-annotated for both molecular parameters and clinical characteristics.
2. Assessment of the prognostic impact and clinical relevance of recurrent gene mutations.
3. Identification of distinct patterns of associations between recurrent mutations with other clinicobiological features in CLL.
4. Perform robust validation of recently proposed prognostication models that incorporate both cytogenetic and molecular lesions as well as immunogenetic characteristics.

Project Start date           1 January 2017

Project Finish date         31 May 2018

Name of sponsor (s) if any

Forms the pilot CLL project within the Harmony alliance; a European Network of Excellence in Hematology and Big Data that is funded through the Innovative Medicines Initiative (IMI)

Contact person (s) for further details:  Lesley Ann Sutton; lesley-ann.sutton@ki.se