Study of the Metabolism in the Lymphatic Niche of CLL

Study of the Metabolic Reprogramming of CLL Cells in Ex-vivo Models of the Lymphatic Niche

Chronic lymphoid leukemia (CLL) is the most common adult leukemia that is characterized by a malignant monoclonal accumulation of tumoral and quiescent B cells in the peripheral blood. In advanced stages of the disease (Binet stage C), this population invades the bone marrow (BM) and proliferate into the lymphoid organs that results in widespread adenopathy. Richter's transformation is a most aggressive serious complication of CLL (transformation of the disease into an aggressive lymphoma) detected based on TEP/CT (Positron Emission Tomography/ computerized tomography) that shows highly derived glucose consumption by cancer cells. Clinical data from CLL patients with disease acutisation showed hypermetabolic lymphadenopathy with high standardized uptake value (SUV) whereas there is low grade tracer uptake into BM. We supposed that the tumor microenvironment of the lymphatic niche promotes the proliferation and glycolytic activity of CLL cells which become particularly resistant to treatment. The development of an ex-vivo tumor model that reproduces the microenvironment of the lymph node niche appears essential to identify and validate new therapeutic targets because despite the therapeutic arsenal available some patients still relapse or are refractory to treatment. Our objectives are to i / Characterize this niche of resistance by the development of an ex-vivo tumor model and ii / Evaluate in-vitro the effectiveness of the association of current treatments (RFC, Ibrutinib or Venetoclax) with anti-metabolic therapies (inhibitors of glycolysis) Our lab is developing an ex-vivo models of the lymphatic niche in CLL based on co-cultures of leukemic cells from patients stimulated with CpG ODN and IL2 with primary human lymphatic fibroblasts (HLF) (EC 12PP15). This co-culture has never been described in the literature and allows us to study the lymphatic niche of CLL patients. Lymph node (LN) exploration in CLL requires invasive access and does not bring any additional information in initial diagnosis. Then, we validated our co-culture model using complementary approaches: increased viability, proliferation, and resistance to Ibrutinib, associated with increased production of anti-apoptotic proteins such as MCL1 and BCL2 after 48 hours of co-culture. Secondly, we studied the metabolism in this resistance niche. We find an increased production of lactate and an acute consumption of glucose, associated with a strong metabolic activation detected by SEAHORSE and by the production of glycolysis enzymes such as hexokinase 2. Our study constitutes an original project because it characterized the energy metabolism of the CLL lymphatic niche by developing an original ex-vivo model and enhanced our understanding of the contribution of the specific microenvironment in the dissociation of metabolic activity using SUV max in BM and lymphatic niche. Anti-metabolic therapies are efficient on co-culture CLL cells and could be an alternative for refractory or relapsed patients under current treatment.

Study Overview

• Status: Recruiting
• Conditions: CLL
• Intervention / Treatment: Other: NO intervention

• Study Type: Observational
• Enrollment (Anticipated): 30

Contacts and Locations

• Study Contact: Name: Caroline Ruetsch-Chelli; Phone Number: 33492039016; Email: ruetsch-chelli.c@chu-nice.fr
• Study Contact Backup: Name: Marcel Deckert

Study Locations

• France
  • Nice, France, 06000
    • Recruiting
    • Nice University Hospital
    • Contact: Caroline Ruetsch-Chelli; Phone Number: 0492039016; Email: ruetsch-chelli.c@chu-nice.fr
    • Principal Investigator: Caroline ruetsch-Chelli

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 90 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

CLL patient

Description

Inclusion Criteria:

• Patients for whom the diagnosis of CLL has been established cytologically and phenotypically (Matutes score)
• Patients over 18 years old
• Patients who have signed the non-objection form
• Untreated patients

Exclusion Criteria:

• Patient with a solid cancer that is progressive or in remission for less than 3 years
• HIV positive patients
• Patients with chronic active hepatitis B or C
• History of allogeneic transplant

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Prospective

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
LLC	Other: NO intervention; No intervention

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Modelisation of a CLL lymphatic niche by creation of an ex-vivo model	Measure of viability and proliferation of CLL cells by annexin PI with multiparameter: flow cytometry device, western blot and validation of the model by testing current therapy (Ibrutinib, anti BTK inhibitor)	48 months
Study of metabolism in CLL ex-vivo model	Anlayse of metabolism (ECAR and OCR) by seahorse methods , PCR qnd protein Multiparameter analyse	48 months
Evaluate in-vitro the effectiveness of the combination of current treatments (RFC, Ibrutinib or Venetoclax) with anti-inflammatory therapies. -metabolic (glycolysis inhibitors)	Viability using multiparameter flow cytometry	48 months

Collaborators and Investigators

• Sponsor: Centre Hospitalier Universitaire de Nice

Study record dates

Study Major Dates

• Study Start (Actual): November 7, 2018
• Primary Completion (Anticipated): December 15, 2022
• Study Completion (Anticipated): December 15, 2022

Study Registration Dates

• First Submitted: November 2, 2022
• First Submitted That Met QC Criteria: November 2, 2022
• First Posted (Actual): November 9, 2022

Study Record Updates

• Last Update Posted (Actual): November 9, 2022
• Last Update Submitted That Met QC Criteria: November 2, 2022
• Last Verified: November 1, 2022

More Information

Terms related to this study

• Other Study ID Numbers: 22Laboimmuno01

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No