Establishment Of Novel Cell Line Research Articles

Establishment of novel cell lines that maintain the features of B cells derived from patients with neuromyelitis optica spectrum disorder

B cells that produce anti–aquaporin-4 (AQP4) antibodies play a crucial role in neuromyelitis optica spectrum disorder (NMOSD) pathogenesis. We previously reported that naïve B (NB) cells from patients with NMOSD, unlike those from healthy controls, exhibit transcriptional changes suggesting the adoption of an antibody-secreting cell (ASCs) phenotype. CD25+ NB cells, whose numbers are increased in NMOSD patients, have a greater capacity to differentiate into ASCs than do CD25- NB cells. Here, we attempted to establish novel B cell subset cell lines from patients with NMOSD to enable molecular analysis of their abnormalities. We generated Epstein-Barr virus–immortalized lymphoblastoid cell lines (LCLs) from CD25+ NB, CD25- NB, and switched memory B (SMB) cells. All LCLs largely maintained the features of the original cell type in terms of cell surface marker expression and could differentiate into ASCs. Notably, CD25+ NB–LCLs derived from patients with NMOSD exhibited a greater capacity to differentiate into SMB-LCLs than did CD25- NB–LCLs derived from patients with NMOSD, suggesting that the established LCLs maintained the characteristics of cells isolated from patients. The LCLs established in this study are likely to be useful for elucidating the mechanism by which cells that produce anti-AQP4 antibodies develop in NMOSD.

Abstract 3915: Establishment of novel cell lines recapitulating the genetic landscape of uveal melanoma and preclinical validation of mTOR as a therapeutic target

Abstract PURPOSE: Uveal melanoma is the most common primary tumor of the eye in adults. There is no standard adjuvant treatment to prevent metastases and no effective therapy in the metastatic setting. Relevant models of uveal melanoma are necessary to preclinical assessment of drugs activity. The goal of this study was to develop cell lines recapitulating the genetic landscape of uveal melanoma and identify therapeutic approaches for treatment using in vitro and in vivo models. EXPERIMENTAL DESIGN: Tumors samples from patients or patient-derived xenografts were put into culture.The cell lines obtained were characterized for chromosomal alterations and reported recurrent mutations. They were tested for sensitivity to a selected panel of drugs including the mTOR inhibitor Everolimus. We assessed mTOR signaling activation in the cell lines and evaluated the effect of Everolimus in patient-derived xenografts. RESULTS: We established seven uveal melanoma cell lines. All display either GNAQ or GNA11 activating mutations, three harbor BAP1 mutations, and one an EIF1AX mutation. No cell line exhibits SF3B1 mutations. mTOR pathway was shown to be activated in these cells. Everolimus reduced viability of the cell lines and significantly delayed in vivo tumor growth of 4 patient-derived xenografts. CONCLUSIONS: Our established panel of uveal melanoma cell lines represents a relevant preclinical tool for the study of this disease. Our in vitro and in vivo data suggest that mTOR inhibition with Everolimus, most probably in combination with other agents, may be considered as a therapeutic option for the management of uveal melanoma. Citation Format: Nabil Amirouchene-Angelozzi, Fariba Nemati, David Gentien, May-Linda Lepage, André Nicolas, Jordan Madic, Amaury Dumont, Guillaume Carita, Jaques Camonis, Laurence Desjardins, Nathalie Cassoux, Sophie Piperno-Neumann, Pascale Mariani, Xavier Sastre, Didier Decaudin, Sergio Roman-Roman. Establishment of novel cell lines recapitulating the genetic landscape of uveal melanoma and preclinical validation of mTOR as a therapeutic target. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3915. doi:10.1158/1538-7445.AM2014-3915

Establishment of novel cell lines recapitulating the genetic landscape of uveal melanoma and preclinical validation of mTOR as a therapeutic target

Uveal melanoma (UM) is the most common primary tumor of the eye in adults. There is no standard adjuvant treatment to prevent metastasis and no effective therapy in the metastatic setting. We have established a unique panel of 7 UM cell lines from either patient's tumors or patient-derived tumor xenografts (PDXs). This panel recapitulates the molecular landscape of the disease in terms of genetic alterations and mutations. All the cell lines display GNAQ or GNA11 activating mutations, and importantly four of them display BAP1 (BRCA1 associated protein-1) deficiency, a hallmark of aggressive disease. The mTOR pathway was shown to be activated in most of the cell lines independent of AKT signaling. mTOR inhibitor Everolimus reduced the viability of UM cell lines and significantly delayed tumor growth in 4 PDXs. Our data suggest that mTOR inhibition with Everolimus, possibly in combination with other agents, may be considered as a therapeutic option for the management of uveal melanoma.

Establishment of novel cell lines latently infected with human immunodeficiency virus 1

Many human immunodeficiency virus 1 (HIV-1) researchers focus on the developing new anti-reservoir therapy to eradicate HIV-1 provirus from the HIV-1-infected patients. HIV-1 provirus is the major obstacle for effective HIV-1 treatment because it integrates into the host genome and can produce a virus progeny after stopping highly active antiretroviral therapy (HAART). We established two novel cell lines latently infected with HIV-1 by limiting dilution cloning of A3.01 cells infected with HIV-1. Analysis of the flanking sequence of HIV-1 proviral DNA integrated into chromosomal cellular DNA revealed that proviral DNA was inserted into different sites of different chromosomes in the two examined cell lines. In these lines, virus reactivation could be induced by a phorbol 12-myristate 13-acetate (PMA) treatment that resulted in a marked increase of the production HIV-1 p24 antigen and appearance of the infectious virus. The novel cell lines latently infected with HIV-1 represent further tool for the study of molecular mechanisms of viral latency and development of anti-reservoir therapy of HIV-1 infection.

Constitutional t(3;11)(p21;q23) in a Family, Including One Member with Lymphoma: Establishment of Novel Cell Lines with this Translocation

We describe a family with an inherited constitutional chromosome translocation (3;11)(p21;q23). Of three proven translocation carriers, one had duodenal malignant lymphoma (B-cell diffuse lymphoma, medium-sized cell type). The t(3;11)(p21;q23) was detected not only in hematopoietic cells including the patient's lymphoma cells, non-pathological bone marrow, and phytohemagglutinin-stimulated peripheral blood, but also in fibroblasts of the skin. We have successfully established an Epstein-Barr virus-transformed B-cell line and a Herpesvirus saimiri-transformed T-cell line from the patient, and found that both cell lines also carried this translocation. The patient's asymptomatic mother and sister had the same chromosomal abnormality. Chromosomal abnormalities of the 11q23 band occur frequently in various hematopoietic malignant disorders, and 3p21 has been linked to the pathogenesis of several solid tumors including carcinomas of the kidney, lung, and breast. Although 11q23 is known to recombine with many different chromosomal segments, t(3;11)(p21;q23) has not been reported to our knowledge. Further assessment is warranted to clarify if this constitutional translocation predisposes to certain malignancies. Our cell lines carrying the novel chromosome translocation would be useful for the molecular analysis of the rearranged genes involving both 3p21 and 11q23.