Single-center, Retrospective Study Aimed to Perform a Molecular Characterization of Aggressive B-cell Lymphomas (Diffuse Large B-cell Lymphoma, High-grade B-cell Lymphoma With MYC and BCL2 and/or BCL6 Translocations, High-grade B-cell Lymphoma Not Otherwise Specified) and to Observe the Clinical Patients' Outcomes According to the 2017 WHO Classification
Conditions: Diffuse Large B Cell Lymphoma (DLBCL)
Sponsor: Andrés José Maria Ferreri

CHARTHER study 
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SINGLE-CENTER, RETROSPECTIVE STUDY AIMED TO PERFORM A MOLECULAR 
CHARACTERIZATION OF AGGRESSIVE B-CELL LYMPHOMAS (DIFFUSE LARGE B-CELL 
LYMPHOMA, HIGH-GRADE B-CELL LYMPHOMA WITH MYC AND BCL2 AND/OR BCL6 
TRANSLOCATIONS, HIGH-GRADE B-CELL LYMPHOMA NOT OTHERWISE SPECIFIED) AND 
TO OBSERVE THE CLINICAL PATIENTS’ OUTCOMES ACCORDING TO THE 2017 WHO 
CLASSIFICATION 
Protocol Acronym: CHARTHER 
Version Number: 1.0 
Date: 21/3/2022 
Sponsor:  IRCCS Ospedale San Raffaele 
Authorized Sponsor Representative:   Fabio Ciceri 
Funding Source:  Roche S.p.A. 
Principal Investigator: Andrés J.M. Ferreri, MD 
Lymphoma Unit, Dept. of Onco-Hematology 
IRCCS Ospedale San Raffaele, Milan, Italy 
Co-Principal Investigator: 
Maurilio Ponzoni, Prof 
Pathology Unit 
IRCCS San Raffaele Scientific Institute, Milan, Italy 
Co- Investigator: 
Lorenza Pecciarini, MS 
Pathology Unit 
IRCCS San Raffaele Scientific Institute, Milan, Italy 
DISCLAIMER 
This material is strictly private, confidential and property of OSR (The Sponsor). Its contents 
must not be disclosed or used except as authorized in writing by the Sponsor. 

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PROTOCOL SIGNATURE PAGE 
Study Title: SINGLE-CENTER, RETROSPECTIVE STUDY AIMED TO PERFORM A 
MOLECULAR CHARACTERIZATION OF AGGRESSIVE B-CELL LYMPHOMAS (DIFFUSE 
LARGE B-CELL LYMPHOMA, HIGH-GRADE B-CELL LYMPHOMA WITH MYC AND BCL2 
AND/OR BCL6 TRANSLOCATIONS, HIGH-GRADE B-CELL LYMPHOMA NOT OTHERWISE 
SPECIFIED) AND TO OBSERVE THE CLINICAL PATIENTS’ OUTCOMES ACCORDING TO THE 
2017 WHO CLASSIFICATION 
Protocol Acronym: CHARTHER 
Protocol Version and Date: Version 1, Date 21/03/2022 
The undersigned has read and understood all the aspects of the protocol detailed within 
this document and agrees to supervise and conduct the trial in accordance with the protocol, 
Guideline for Good Clinical Practice ICH E6 (R2), Declaration of Helsinki, and all applicable 
regulatory requirements. 
Principal 
Investigator 
Andrés J.M. Ferreri 
Signature 
Affiliation 
Date 
Authorized 
Sponsor  
Fabio Ciceri 
Signature 
Affiliation 
Date 

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Table of contents 
KEY TRIAL CONTACTS .................................................................................................... 4 
SYNOPSIS (English) ......................................................................................................... 5 
ABBREVIATIONS AND DEFINITIONS .............................................................................. 8 
Abbreviations ................................................................................................................... 8 
Definitions ............................................................... Errore. Il segnalibro non è definito. 
BACKGROUND .................................................................................................................. 8 
RATIONALE ....................................................................................................................... 9 
OBJECTIVES ................................................................................................................... 10 
THERAPEUTIC OUTCOME .......................................................................................... 10 
STUDY DESIGN ............................................................................................................... 11 
Study duration  .............................................................................................................. 11 
STUDY POPULATION ..................................................................................................... 11 
METHODS ........................................................................................................................ 12 
Treatments .................................................................................................................... 12 
Recruitment and Enrollment ................................... Errore. Il segnalibro non è definito. 
Assignment of Subject Identification .............................................................................. 12 
Data Collection .............................................................................................................. 12 
Histopathology and Immunohistochemistry ................................................................... 12 
FISH .............................................................................................................................. 13 
STUDY VARIABLES ........................................................................................................ 14 
DATA MANAGEMENT ..................................................................................................... 14 
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN ........................................... 15 
ETHICAL CONSIDERATIONS AND INFORMED CONSENT .......................................... 15 
PATIENT SAFETY ........................................................................................................... 16 
FINANCE AND INSURANCE ........................................................................................... 16 
OWNERSHIP OF DATA AND PUBLICATIONS .............................................................. 16 
REFERENCES ................................................................................................................. 16 

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KEY TRIAL CONTACTS 
Sponsor 
IRCCS Ospedale San Raffaele 
Via Olgettina, 60 
20132 – Milano, Italy 
Authorized Sponsor 
Representative 
Fabio Ciceri, Prof/MD  
Head of Onco-Hematology and Bone Marrow Transplant Unit 
IRCCS Ospedale San Raffaele, Milan, Italy 
Via Olgettina 60  
20132 – Milan, Italy 
Email: ciceri.clinicaltrials@hsr.it  
Principal 
Investigator 
Andrés J.M. Ferreri, MD 
Lymphoma Unit, Dept. of Onco-Hematology 
IRCCS Ospedale San Raffaele, Milan, Italy 
Via Olgettina 60  
20132 – Milan, Italy 
Telephone: 0039-02-2643 7649 
E-mail: ferreri.andres@hsr.it
Co-Investigators 
Maurilio Ponzoni, Prof 
Pathology Unit 
IRCCS San Raffaele Scientific Institute, Milan, Italy 
Via Olgettina 60  
20132 – Milan, Italy 
Telephone: 0039-02-2643 2544 
E-mail: ponzoni.maurilio@hsr.it
Lorenza Pecciarini, MS 
Pathology Unit 
IRCCS San Raffaele Scientific Institute, Milan, Italy 
Via Olgettina 60  
20132 – Milan, Italy 
Telephone: 0039-02-2643 2512 
E-mail: pecciarini.lorenza@hsr.it
Trial Office 
Ufficio Ricerche Cliniche – Stem Cell Program 
Coordinamento 
  Stefania Trinca 
Regolatorio         
   Dott. Vincenzo Mercurio 
Funding source(s) 
Roche S.p.A.  

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SYNOPSIS (English) 
Study Title 
Single-center, retrospective study aimed to perform a molecular 
characterization of aggressive b-cell lymphomas (diffuse large b-cell 
lymphoma, high-grade B-cell lymphoma with MYC and BCL2 and/or 
BCL6 translocations, high-grade B-cell lymphoma not otherwise 
specified) and to observe the clinical patients’ outcomes according 
to the 2017 WHO classification 
Protocol 
Acronym/Number 
CHARTHER 
Protocol Version 
and Date 
1.0  - 21 March 2022
Sponsor 
IRCCS Ospedale San Raffaele 
Via Olgettina 60 – 20132, Milan, Italy 
Principal 
Investigator 
Andrés J.M. Ferreri, MD 
Lymphoma Unit, Dept. of Onco-Hematology 
IRCCS Ospedale San Raffaele 
Via Olgettina 60  
20132 – Milan, Italy 
Telephone: 0039-02-2643 7649 
E-mail: ferreri.andres@hsr.it
Study 
Description 
This is a retrospective analysis of the presence of chromosomal 
rearrangements involving MYC, BCL2 and BCL6 genes by FISH in 
a retrospective, single-center series of large B-cell lymphomas, in 
order to allow a better classifications of these cases according to the 
current WHO 2017 classification. If this evaluation will be confirmed 
as a reliable method to reclassify in different groups the evaluated 
tumors, it can be subsequently routinely apply to indicate the best 
treatment approaches in this high-risk setting of patients with a 
significant impact on patients’ morbidity and mortality, and also on 
the health system and related costs. 
Primary 
Secondary 
Objectives 
1. To re-classify retrospective
series of large B-cell
lymphomas (LBCL) according
to the current WHO 2017
classification by using FISH for
the detection of MYC, BCL2
and BCL6 rearrangements.
2. To establish the association
between re-classification of
LBCL using FISH for the
detection of MYC, BCL2 and
1. To establish the association
between re-classification of
LBCL using FISH for the
detection of MYC, BCL2 and
BCL6 rearrangements and
patients’ outcomes.
2. To establish the association
between re-classification of
LBCL and risk of relapse.
3. To demonstrate that re-
classification of LBCL exploiting
FISH for the detection of MYC,

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BCL6 rearrangements and 
response to therapy. 
BCL2 and BCL6 rearrangements 
is a reliable tool to support 
treatment decision plan. 
4. To complete IHC analysis for 
further and/or novel pattern of 
antigen expression in HGBL. 
5. To  estimate  the  prevalence 
of MYC, BCL2 and BCL6 
rearrangements/ amplifications 
detected by FISH in LBCL 
Study Population Adult subjects (> 18 years old) of any age and gender affected by 
large B-cell lymphoma. We expect to enroll around 555 subjects.  
Inclusion Criteria 
• Histologically-confirmed diagnosis of LBCL (i.e., DLBCL, HGBCL, 
unclassifiable B-cell lymphoma with features intermediate between 
DLBCL and BL), or aggressive B-cell lymphomas with blastoid 
morphology (excluding blastoid variant of mantle cell lymphoma 
and lymphoblastic lymphoma) 
• Histopathological diagnosis performed between 2000 and 2019.  
• HIV sero-negativity 
• Age ≥18 
• Treatment with R-CHOP or other intensified polychemotherapy 
regimen 
• Availability of adequate histopathological samples at diagnosis for 
FISH analysis 
• Availability of clinical records and outcomes data 
 
Exclusion 
Criteria 
• Histologic diagnosis other than DLBCL or unclassifiable B-cell 
lymphoma with features intermediate between DLBCL and BL 
Statistical Design 
Subjects who fulfil selection criteria for enrolment into the study will 
undergo 
local 
histopathological 
revision 
and 
analysis 
of 
chromosomal rearrangements involving MYC, BCL2 and BCL6 
genes by FISH. Moreover, completion of IHC analysis for further 
and/or novel pattern of antigen expression in HGBL will be 
performed. 
 
Histopathology and Immunohistochemistry 
Nodal and extra nodal tissues with a diagnosis of DLBCL, 
lymphomas with intermediate features between Burkitt lymphoma 
and DLBCL, Burkitt-like lymphomas, and atypical Burkitt lymphomas 
will be searched within the files of Pathology Unit at San Raffaele 
Scientific Institute. The selection process will rely on sequential 
steps. First, we will retrieve archival cases from our tissue repository 
placed in a remote place outside our Institution. Second, we will 
carefully consider only tissue blocks showing enough material to 
allow completion of immunohistochemistry and FISH analyses. 

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Third, original slides will be reviewed and re-classified according to 
present WHO (2017) classification of lymphomas. Given that it is 
expected that a brand-new Classification will be shortly released in 
the forthcoming months by WHO, it is likely that additional diagnostic 
criteria will be further included within the central pathology review.  
Each DLBCL case will be stained, whenever not previously 
evaluated, with CD10, bcl-6 and MUM-1 antibodies and their 
respective 
secondary 
antibodies 
using 
an 
automated 
immunostainer. Staining data will be interpreted semiquantitatively. 
According to immunohistochemical results, each case will be further 
classified following to Hans’ algorithm criteria. Each case will be also 
evaluated for c-MYC immunoreactivity and will be considered 
positive whenever more than 40% of cells will be stained for this 
marker. The correlation with FISH results will enable to select the 
actual high grade B cell lymphomas, which will be matched with 
clinical parameters and therapeutic responses.  
 
 
FISH  
Interphase Fluorescence In Situ Hybridization (I-FISH) for detection 
of MYC, BCL2 and BCL6 rearrangements will be performed on the 
previously histologically reviewed large B-cell lymphoma cases. A 
“split-apart” FISH approach will be used: the chosen probes for each 
gene of interest consist of two direct labelled gene locus regions 
(green and orange), hybridizing distal and proximal to the gene 
breakpoint and separation/fusion of the two signals will indicate 
presence/absence of the rearrangements. Three 4 μm tissue 
sections will prepared for each sample, they will be pretreated 
according to the ZytoLight FISH-Tissue Implementation Kit protocol 
(Zytovision, GmbH). Hybridization will be then carried out overnight 
on a Thermobrite hybridizer (Euroclone), using the ZytoLight SPEC 
MYC Dual Color Break Apart Probe, ZytoLight SPEC BCL2 Dual 
Color Break Apart Probe, ZytoLight SPEC BCL6 Dual Color Break 
Apart Probe, following the manufacturer protocol; the next day, after 
unbound probes are removed by stringency washing steps, the 
slides will be counterstained with DAPI. Hybridized probes will be 
visualized using a fluorescence microscope (AxioImager Z2, Zeiss) 
equipped with excitation and emission filters specific for the 
fluorochromes by which the FISH probes are directly labeled. FISH 
semiautomatic analysis will be performed using the Metafer4 
software (Metasystems): FISH signals will be counted in 200 
interphase nuclei from at least 4 to 8 areas selected for well-
preserved cellular and nuclear morphology to ensure representative 
sampling and to avoid nuclear truncation artifacts; a signal split, 
which indicates gene rearrangement, will be counted when the 
distance between splitted green and orange signals is ≥2 of the 
estimated signal diameter. The already defined laboratory cutoff for 
gene rearrangements will be used (2% for all the used probes). 

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Results will be summarized for each case and reported to the 
pathologist. 
Planned 
Study 
Periods  
Duration of the Enrollment period: 1 year 
Duration of the retrospective Follow-up: NA 
Duration of total study period: 1 year 
 
 
ABBREVIATIONS AND DEFINITIONS 
Abbreviations 
BL 
Burkitt lymphoma 
CR 
complete remission 
DLBCL 
diffuse large B-cell lymphoma 
DOR 
Duration of response 
FISH 
fluorescent in situ hybridization 
HGBL 
High-grade B-cell Lymphoma 
IC 
Informed Consent 
I-FISH 
Interphase Fluorescence In Situ Hybridization 
IHC 
immunohistochemistry  
OS 
Overall Survival  
PFS 
Progression-free Survival 
PR 
partial response 
 
 
BACKGROUND 
The 2017 WHO classification introduced the new clinical entity “High-grade B-cell 
Lymphoma” (HGBL), including large B-cell lymphomas with chromosomal rearrangements 
involving MYC gene (region 8q24) and BCL2 gene (region 18q21) (so called “double hit 
lymphoma”, representing 60% of HGBL) and/or BCL6 (region 3q27) (also called “triple hit 
lymphoma”, representing 20% of HGBL), identified by means of fluorescent in situ 
hybridization (FISH) and/or conventional cytogenetic analysis. HGBL replaced the old WHO 
2008 category of ‘unclassifiable B-cell lymphoma with features intermediate between diffuse 
large B-cell lymphoma and BL’ and encompasses also diffuse large B-cell lymphoma 
(DLBCL) by morphology harboring MYC and BCL-2 and/or BCL-6 rearrangements. 
Clinically, HGBL are characterized by adverse prognosis and poor response to standard 
treatments, such as R-CHOP regimen, with a median OS of 12 months or less. The role of 

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intensified chemo-immunotherapy approaches has been investigated. Promising results 
emerged when treating patients with R-DA-EPOCH regimen, together with adequate CNS 
prophylaxis. [Petrich AM, Blood 2014]. Excellent efficacy and safety results were also 
reported in a multicenter phase II trial addressing a dose-dense, short-term therapy in HIV-
positive patients diagnosed with high-risk BL or HGBL with MYC rearrangements [Ferreri et 
al. BJH 2021]. In the last decades, the “CARMEN” regimen was adopted at out institution 
and at several Italian Centers also for the treatment of HIV-negative patients diagnosed with 
high-risk DLBCL as well as B-cell lymphoma unclassifiable with features intermediate 
between DLBCL and BL with alterations of c-MYC. A retrospective series of 63 HIV-negative 
and –positive patients (22 HGBL) confirmed good tolerability and efficacy: after induction, 
18 (82%) HGBL patients achieved a response, which was CR in 10 (45%) patients. At a 
median follow-up of 54 (2-131) months, the 5-year PFS and OS among HGBL were 67% 
and 66%, respectively. HIV seropositivity did not modify outcome. 
Although FISH represents the gold standard for detection of MYC, BCL2 and BCL6 
rearrangements, it is not routinely recommended in all cases at diagnosis. Indeed, a 
consensus has not yet been reached to provide specific guidelines as to which LBCL should 
be evaluated. Some believe that all DLBCL should have genetic studies for the detection of 
the three genes rearrangements, whereas others would limit them, for example, to cases 
with a GCB phenotype and/or high-grade morphology or to cases with >40% MYC+ cells in 
immunohistochemistry (IHC). In particular, at our institution, FISH for MYC is considered 
when MYC expression by IHC is >40%, and FISH for BCL-2 and BCL-6 are performed only 
when FISH for MYC is positive. However, since the presence MYC and BCL2 and/or BCL6 
rearrangements is not invariably associated with IHC overexpression, the current diagnostic 
approach could have underestimated the real incidence of HGBL leading to the 
undertreatment of these patients. 
 
RATIONALE 
We propose to retrospectively analyze the presence of chromosomal rearrangements 
involving MYC, BCL2 and BCL6 genes by FISH in a retrospective, single-center series of 
large B-cell lymphomas, in order to allow a better classifications of these cases according 
to the current WHO 2017 classification. If this evaluation will be confirmed as a reliable 
method to reclassify in different groups the evaluated tumors, it can be subsequently 

CHARTHER study 
 
 
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routinely apply to indicate the best treatment approaches in this high-risk setting of patients 
with a significant impact on patients’ morbidity and mortality, and also on the health system 
and related costs. Moreover, this study could give important information about the real 
incidence and predictive value of MYC, BCL2 and BCL6 rearrangements. 
 
OBJECTIVES 
Primary objectives:  
1. To re-classify retrospective series of large B-cell lymphomas (LBCL) according to the 
current WHO 2017 classification by using FISH for the detection of MYC, BCL2 and 
BCL6 rearrangements. 
2. To establish the association between re-classification of LBCL using FISH for the 
detection of MYC, BCL2 and BCL6 rearrangements and survival. 
 
Secondary objectives:  
1. To establish the association between re-classification of LBCL using FISH for the 
detection of MYC, BCL2 and BCL6 rearrangements and patients’ response to 
treatments.  
2. To establish the association between re-classification of LBCL and risk of relapse.  
3. To demonstrate that re-classification of LBCL exploiting FISH for the detection of 
MYC, BCL2 and BCL6 rearrangements is a reliable tool to support treatment decision 
plan. 
4. To complete IHC analysis for further and/or novel pattern of antigen expression in 
HGBL. 
5. To  estimate  the  prevalence of MYC, BCL2 and BCL6 rearrangements/ 
amplifications detected by FISH in LBCL 
6. To describe the main treatment patterns according to disease characteristics. 
 
THERAPEUTIC OUTCOME 
 
Primary outcome:  
1. Overall Survival (OS) 
 

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Secondary outcome: 
1. Progression-free Survival (PFS) 
2. Overall Response Rate (ORR; partial response [PR] + complete remission [CR]) 
3. Duration of response (DOR; partial response [PR] + complete remission [CR]) 
4. Relapse rates and patterns 
5. Incidence of MYC, BCL2 and BCL6 rearrangements in LBCL 
 
STUDY DESIGN 
Single 
center, 
retrospective 
observational, 
pharmacological 
and 
of 
molecular 
characterization study. 
Study duration 1 year 
 
STUDY POPULATION  
Adult subjects (> 18 years old) of any age and gender affected by large B-cell lymphomas. 
 
Inclusion Criteria:  
• Histologically-confirmed diagnosis of LBCL (i.e., DLBCL, HGBCL, unclassifiable B-cell 
lymphoma with features intermediate between DLBCL and BL), or aggressive B-cell 
lymphomas with blastoid morphology (excluding blastoid variant of mantle cell 
lymphoma and lymphoblastic lymphoma) 
• Histopathological diagnosis performed between 2000 and 2019. 
 
• HIV sero-negativity 
• Age ≥18 
• Treatment with R-CHOP or other intensified polychemotherapy regimen 
• Availability of adequate histopathological samples at diagnosis for FISH analysis 
• Availability of clinical records and outcomes data 
 
Exclusion criteria 
• Histologic diagnosis other than DLBCL or unclassifiable B-cell lymphoma with features 
intermediate between DLBCL and BL 
 

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METHODS 
Diagnostic tissue samples of patients who fulfil selection criteria for enrolment into the study 
will undergo local histopathological revision and analysis of chromosomal rearrangements 
involving MYC, BCL2 and BCL6 genes by FISH. Moreover, completion of IHC analysis for 
further and/or novel pattern of antigen expression in HGBL will be performed. 
 
Treatments: anthracycline-based combinations followed or not by involved-field 
radiotherapy 
 
 
Assignment of Subject Identification 
The participants will be assigned a unique trial specific number and will henceforth be 
identified by that number in the database. 
 
Data Collection 
The data of all patients enrolled in the study will be collected from clinical folders and source 
documents (e.g., hospital records; clinical and office charts; laboratory notes etc.) and 
registered in the database as described in the following paragraphs. 
 
Histopathology and Immunohistochemistry 
Nodal and extra nodal tissues with a diagnosis of DLBCL, lymphomas with intermediate 
features between Burkitt lymphoma and DLBCL, Burkitt-like lymphomas, and atypical 
Burkitt lymphomas will be searched within the files of Pathology Unit at San Raffaele 
Scientific Institute. The selection process will rely on sequential steps. First, we will retrieve 
archival cases from our tissue repository placed in the Pathology unit of the OSR. Second, 
we will carefully consider only tissue blocks showing enough material to allow completion 
of immunohistochemistry and FISH analyses. Third, original slides will be reviewed and re-
classified according to present WHO (2017) classification of lymphomas. Given that it is 
expected that a brand-new Classification will be shortly released in the forthcoming months 
by WHO, it is likely that additional diagnostic criteria will be further included within the central 
pathology review.  

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Each DLBCL case will be stained, whenever not previously evaluated, with CD10, bcl-6 and 
MUM-1 antibodies and their respective secondary antibodies using an automated 
immunostainer. Staining data will be interpreted semiquantitatively. According to 
immunohistochemical results, each case will be further classified following to Hans’ 
algorithm criteria. Each case will be also evaluated for c-MYC immunoreactivity and will be 
considered positive whenever more than 40% of cells will be stained for this marker. The 
correlation with FISH results will enable to select the actual high grade B cell lymphomas, 
which will be matched with clinical parameters and therapeutic responses.  
 
FISH  
Interphase Fluorescence In Situ Hybridization (I-FISH) for detection of MYC, BCL2 and 
BCL6 rearrangements will be performed on the previously histologically reviewed large B-
cell lymphoma cases. A “split-apart” FISH approach will be used: the chosen probes for 
each gene of interest consist of two direct labelled gene locus regions (green and orange), 
hybridizing distal and proximal to the gene breakpoint and separation/fusion of the two 
signals will indicate presence/absence of the rearrangements. Three 4 μm tissue sections 
will prepared for each sample, they will be pretreated according to the ZytoLight FISH-
Tissue Implementation Kit protocol (Zytovision, GmbH). Hybridization will be then carried 
out overnight on a Thermobrite hybridizer (Euroclone), using the ZytoLight SPEC MYC Dual 
Color Break Apart Probe, ZytoLight SPEC BCL2 Dual Color Break Apart Probe, ZytoLight 
SPEC BCL6 Dual Color Break Apart Probe, following the manufacturer protocol; the next 
day, after unbound probes are removed by stringency washing steps, the slides will be 
counterstained with DAPI. Hybridized probes will be visualized using a fluorescence 
microscope (AxioImager Z2, Zeiss) equipped with excitation and emission filters specific for 
the fluorochromes by which the FISH probes are directly labeled. FISH semiautomatic 
analysis will be performed using the Metafer4 software (Metasystems): FISH signals will be 
counted in 200 interphase nuclei from at least 4 to 8 areas selected for well-preserved 
cellular and nuclear morphology to ensure representative sampling and to avoid nuclear 
truncation artifacts; a signal split, which indicates gene rearrangement, will be counted when 
the distance between splitted green and orange signals is ≥2 of the estimated signal 
diameter. The already defined laboratory cutoff for gene rearrangements will be used (2% 

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for all the used probes). Results will be summarized for each case and reported to the 
pathologist. 
STUDY VARIABLES 
Conventional anagraphic variables, findings of histopathological diagnosis, results of IHC 
and FISH analyses, staging, therapeutic management and outcome will be considered.  
DATA MANAGEMENT 
Data Recording and Record-Keeping 
All relevant data collected during the study for all of the patients enrolled in the study shall 
be entered in a database (password protected excel) from source documents (e.g., hospital 
records; clinical and office charts; laboratory notes etc.) by the responsible investigator or 
someone authorized by the investigator in a timely manner (as soon as possible after the 
information is collected) to ensure that they are clear, complete and correct.  
On all trial-specific documents, other than the signed consent, the participant will be referred 
to by the trial participant number/code, not by name.  
The investigator shall arrange for the retention of patient files, other source data, and the 
Trial Master File for at least 7 years after the completion or discontinuation of the study. 
 
Data Protection 
The confidentiality of participant data, including, but not limited to forms, records and 
samples and participant privacy will be maintained according to the GDPR (UE) 2016/679. 
The trial staff will ensure that the participants’ anonymity is maintained. The participants will 
be identified only by initials and a Patient ID number on the ICF and the electronic database. 
All documents will be stored securely and only accessible by trial staff and authorized 
personnel. 
 
Data anonymization 
Since this is a retrospective study without ad hoc informed consent, in order to ensure 
anonymity, the following procedure will be followed for all data of all subjects: 
the assigned patient ID will not be related to the identification code present in the patient's 
clinical material; this will be guaranteed with the following procedures: 
- 
A patient code will be associated with the clinical material (nominal); 

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- 
The code will be transcribed in a second document and associated with a second 
code obtained by randomization (called "Patient ID"); 
- 
The patient ID will then be reported in the CRF and the intermediate document will 
be destroyed at termination of data collection. 
At the end of this process and in the continuation of the study, patient data will be treated 
completely anonymously. 
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN 
Clinical characteristics and response rates of the therapeutic subgroups will be compared 
using the α2 test or Fisher exact test for categorical variables, according to the sample size. 
Factors potentially conditioning outcome will be assessed by α2 test and logistic regression.  
Survival curves will be generated by the Kaplan-Meier method. Overall survival (OS) will be 
calculated from the date of pathologic diagnosis to death or to the last date of follow-up, 
while progression-free survival (PFS) will be calculated from the first day of treatment to 
relapse, progression or death, or to the last date of follow-up. Survival rates will be reported 
as 2-year OS and 95% CI. Impact on survival of clinical and therapeutic variables will be 
evaluated by comparing the entire OS curves by means of the log-rank test.  
The independent prognostic value of variables will be analyzed using the Cox model. The 
year of diagnosis will be included as a continuous variable in the multivariate analyses to 
exclude the possibility that modern medical management could influence outcome more 
than the treatment itself. Backward stepwise regression will be performed to identify the 
most powerful predictors of survival. All the probability values will be two-sided. Statistical 
analysis will be performed with Statistical software (Statistica 10.0, Tuxon, AZ, USA) on a 
unique database including clinical and pathological data on an estimated sample size of 
555 patients. 
ETHICAL CONSIDERATIONS AND INFORMED CONSENT 
The Investigator and all study staff will ensure that the clinical study is conducted in 
accordance with good clinical practice and institutional requirements, including those for 
subject privacy, Ethics Committee approval and record retention, and with the Declaration 
of Helsinki (revised October 2008) and in compliance with principles defined in the 
Determinazione of the Agenzia Italiana del Farmaco, dated 20 March 2008. The protocol, 
being retrospective, will be notified to the IEC of the IRCCS San Raffaele Scientific Institute 

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of Milan. In the context of clinical practice, patients already provided consent to use of health 
data for research purposes. 
PATIENT SAFETY 
Given the observational retrospective nature of the study, for the patients enrolled in this 
study, there are no additional physical, psychological, or social adverse effects compared 
to the standard of care clinical practice. 
FINANCE AND INSURANCE 
The study is non-profit in nature.  
Funding for the study has been received from Roche S.p.A.  
This funding will cover the whole costs of the study.  
Given the observational retrospective nature of this study, no insurance policy is needed. 
OWNERSHIP OF DATA AND PUBLICATIONS 
The Sponsor has the ownership of all data and results collected during the study. The 
results of this study will be presented anonymously to national and international scientific 
meetings and reported in scientific publication focused on translational research. 
Each publication of protocol results will occur in mutual agreement between the principal 
investigator and any other investigators involved. All data will be treated in confidence by 
the principal investigator and all others involved in the protocol until publication. Final results 
may only be published (orally or in writing) with agreement from the principal investigator. 
This is essential for a thorough exchange of information between theaforementioned parties 
and will ensure that the opinions of all parties involved have been heard before publication. 
This agreement, which does not include any veto right or right of censorship for any of the 
parties involved, may not be refused without good reason. 
The Principal Investigator/Sponsor is responsible for the preparation of an annual report on 
the clinical study to be sent to the Ethics Committee and for the preparation of the final 
report of the clinical study. 
 
REFERENCES 
1. Ferreri AJM, Cattaneo C, Lleshi A, et al. A dose-dense short-term therapy for human 
immunodeficiency virus/acquired immunodeficiency syndrome patients with high-risk 

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Burkitt lymphoma or high-grade B-cell lymphoma: safety and efficacy results of the 
"CARMEN" phase II trial. Br J Haematol. 2021;192(1):119-128.  
2. Petrich AM, Gandhi M, Jovanovic B, et al. Impact of induction regimen and stem cell 
transplantation on outcomes in double-hit lymphoma: a multicenter retrospective 
analysis. Blood. 2014;124(15):2354-2361.