This document summarizes research on generating stable monoclonal cell lines from a chronic lymphocytic leukemia (CLL) mouse model. The cell lines, called EMC lines, exhibit constant Btk and Akt signaling that is dependent on B-cell receptor signaling, mimicking human CLL. The EMC lines provide a tool to study CLL cell biology and test novel targeted therapies in vitro and in vivo, such as the Btk inhibitor ibrutinib. When engrafted into mice, the EMC lines allow rapid evaluation of drug treatment on CLL progression within reasonable timeframes.

1. PRESENTED BY
AYUSH JAIN
ROLL NO: 16
DEPARTMENT OF BIOTECNOLOGY
V.N.S.G.U

3. Abstract
In vitro survival and proliferation of CLL is depend on phosphorylation of Burton's tyrosine
kinase (Btk) and Akt(protein kinase).
To reduce cell viability and proliferation, cells were treated with small molecule inhibitors
specific for Btk (ibrutinib) or PI3K (idelalisib).

4. Introduction
Chronic lymphocytic leukaemia (CLL) is a type of blood and bone marrow cancer
characterized by accumulation of mature B cells.
Evidence supports key role of B-cell receptor (BCR) signalling in CLL pathogenesis.
CLL have higher basal, cell-autonomous Ca2+ signaling, dependent on an internal BCR epitope.

5. Objective
To generated stable monoclonal cell lines from a CLL mouse model that exhibits constitutive
Btk and Akt signaling.
To study CLL responds to specific inhibitors for novel targeted therapies.

6. (A)Generation and characterization of cell line from IgH. TEμ
CLL mice

7. B) EMC cell lines provide a novel in vitro tool to test therapeutic
drugs for CLL

8. C) Engraftment of EMC cell lines into Rag1–/– mice as a tool to study
novel CLL therapeutics

9. EMC4-engrafted mice were receive either ibrutinib or vehicle to
investigated, if ibrutinib treatment affected disease progression.

10. Discussion
These cell lines can be cultured In vitro (EMC2, EMC4 and EMC6) for long periods of time
and transferred into mice, thus providing a platform to study BCR signaling in CLL and to
investigate the efficacy of small molecule inhibitors.
The efficacy of the novel CLL therapeutics ibrutinib and idelalisib is not only based on their
effects on BCR-mediated survival and proliferation, but also on cellular adhesion and migration
in the context of the CLL microenvironment.

11. Conclusion
The rapid CLL development in engrafted mice facilitates the evaluation of therapeutic strategies
within reasonable time schedules and contrasts with the slow disease development in CLL
mouse models.
EMC cell lines thus provide a novel preclinical platform to study CLL cell biology and to test
efficacy of novel targeted therapy combinations.

12. REFFERNCES
Singh, S. P., Pillai, S. Y., de Bruijn, M. J., Stadhouders, R., Corneth, O. B., van den Ham, H. J.,
& Spaargaren, M. (2017). Cell lines generated from a chronic lymphocytic leukemia mouse
model exhibit constitutive Btk and Akt signaling. Oncotarget, 8(42), 71981.