Molecular characterization of neoplastic and normal “sister” lymphoblastoid B-cell lines from chronic lymphocytic leukemia

Myhrinder, Anna Lanemo; Hellqvist, Eva; Bergh, Ann-Charlotte; Jansson, Mattias; Nilsson, Kenneth; Hultman, Per; Jonasson, Jon; Buhl, Anne Mette; Pedersen, Lone Bredo; Jurlander, Jesper; Klein, Eva; Weit, Nicole; Herling, Marco; Rosenquist, Richard; Rosén, Anders

doi:10.3109/10428194.2013.764418

Cited by 15 publications

(16 citation statements)

References 39 publications

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“…This is in line with the fact that the aggressiveness of the cancer cells correlates with increasing SA levels [18, 19]. HG3 and CI have an unmutated immunoglobulin heavy chain variable ( IGHV ) gene, which is associated with high cell proliferation and a more aggressive form of CLL compared with Wa-osel and AIII cells, which have a mutated IGHV gene, a signature of less aggressive indolent CLL cells [17]. …”

Section: Discussionmentioning

confidence: 62%

“…SA plays an important role in a variety of biological processes in cells. In this study, we were primarily interested to verify any correlations with the aggressiveness of the CLL cell lines and SA expression levels [17]. …”

Section: Discussionmentioning

confidence: 99%

“…Four CLL cell lines were used, HG3, CI, Wa-osel, and AIII [17]. The cell lines were cultured in RPMI-1640 medium (Invitrogen, San Diego, CA, USA) supplemented with 10 % fetal bovine serum (FBS, Invitrogen) and 50 μg/mL gentamycin (Invitrogen) and incubated in 37 °C with 5 % CO 2 in 100 % humidity.…”

Section: Methodsmentioning

confidence: 99%

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Different expression levels of glycans on leukemic cells—a novel screening method with molecularly imprinted polymers (MIP) targeting sialic acid

et al. 2016

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Sialic acid (SA) is normally expressed on the cell membranes and is located at the terminal position of the sugar chains. SA plays an important role for regulation of the innate immunity, function as markers of the cells and can be recognized by a variety of receptors. Interestingly, the level of SA expression is increased on metastatic cancer cells. The availability of specific antibodies against SA is limited and, therefore, biomarker tools for detection of SA are lacking. We have recently presented a novel method for specific fluorescence labeling of SA molecular imprinted polymers (MIP). Here, we have performed an extended screening of SA expression by using SA-MIP and included four different chronic lymphocytic leukemia (CLL) cell lines, conveniently analyzed by flow cytometry and fluorescence microscopy. SA expression was detected in four cell lines at different levels, and the SA expression were verified with lectin-FITC. These results show that SA-MIP can be used as a plastic antibody for detection of SA using both flow cytometry and fluorescence microscopy. We suggest that SA-MIP can be used for screening of different tumor cells of various stages, including CLL cells.

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Section: Discussionmentioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

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Different expression levels of glycans on leukemic cells—a novel screening method with molecularly imprinted polymers (MIP) targeting sialic acid

et al. 2016

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“…To study if TP53 disruption directly affects BCR signaling, we employed three cell lines: HG3 and PGA1, which are CD5+/CD19+ CLL lymphoblastoid cell lines and PG-EBV, which is an EBV transformed B-cell lymphoblastoid cell line, all of which are TP53 unmutated as verified by us through sequencing(34). Over a dose range of 0.25 μM to 5 μM of ibrutinib the HG3 and PGA1 lines were not sensitive to ibrutinib while the PG-EBV line displayed an IC50 value of ~2.5 μM (Figure 6A; Supplementary Figure 5A; Supplementary Figure 6A).…”

Section: Resultsmentioning

confidence: 99%

Cell-Intrinsic Determinants of Ibrutinib-Induced Apoptosis in Chronic Lymphocytic Leukemia

Amin

Balasubramanian

Saiya-Cork

et al. 2017

Clinical Cancer Research

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Purpose Ibrutinib, a Bruton’s tyrosine kinase (BTK) inhibitor, is approved for the treatment of relapsed CLL and CLL with del17p. Mechanistically, ibrutinib interferes with BCR signaling as well as multiple CLL cell to microenvironment interactions. Given the importance of ibrutinib in the management of CLL, a deeper understanding of factors governing sensitivity and resistance is warranted. Experimental Design We studied 48 longitudinally sampled paired CLL samples, 42 of which were procured before and after standard CLL chemotherapies, and characterized them for well-studied CLL molecular traits as well as by whole exome sequencing and SNP 6.0 array profiling. We exposed these samples to 0.25 μM – 5 μM of ibrutinib ex vivo and measured apoptosis fractions as well as BCR signaling by immunoblotting. We disrupted TP53 in HG3, PGA1 and PG-EBV cell lines and measured BCR signaling and ibrutinib responses. Results CLL samples demonstrated a surprisingly wide range of ex vivo sensitivities to ibrutinib with IC50 values ranging from 0.4 μM – 9.7 μM. Unmutated IGVH status, elevated ZAP70 expression and trisomy 12 were associated with heightened sensitivity to ibrutinib treatment. Five CLL samples were substantially more resistant to ibrutinib following relapse from chemotherapy; of these, three had acquired a del17p/TP53 mutated status. A validation sample of 15 CLL carrying TP53 mutations, of which 13 carried both del17p and a TP53 mutation confirmed substantially less sensitivity to ibrutinib-induced apoptosis. Conclusions This study identifies that CLL harboring del17p/TP53 mutated cells are substantially less sensitive to ibrutinib-induced apoptosis than del17p/TP53 wild type cells.

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“…Only original stock propagated immediately upon arrival for 2 to 3 passages was picked for studies and cultures terminated after the 10th round of passaging (5-7 weeks). Upon thawing for experimentation between 2012 and 2015, all lines were authenticated by flow cytometry confirming their characteristic immunophenotype, including Ig composition and TCL1 levels (28)(29)(30). The same applied to the FL/DLBCL lines LP, FN, CJ, LR, MS, DBr, DS, EJ established and provided by Dr. R.J. Ford (UT M.D.…”

Section: Cell Isolation and Culturesmentioning

confidence: 99%

A Novel Recombinant Anti-CD22 Immunokinase Delivers Proapoptotic Activity of Death-Associated Protein Kinase (DAPK) and Mediates Cytotoxicity in Neoplastic B Cells

Lilienthal

Lohmann

Crispatzu

et al. 2016

Molecular Cancer Therapeutics

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The serine/threonine death-associated protein kinases (DAPK) provide pro-death signals in response to (oncogenic) cellular stresses. Lost DAPK expression due to (epi)genetic silencing is found in a broad spectrum of cancers. Within B-cell lymphomas, deficiency of the prototypic family member DAPK1 represents a predisposing or early tumorigenic lesion and high-frequency promoter methylation marks more aggressive diseases. On the basis of protein studies and meta-analyzed gene expression profiling data, we show here that within the low-level context of B-lymphocytic DAPK, particularly CLL cells have lost DAPK1 expression. To target this potential vulnerability, we conceptualized B-cell-specific cytotoxic reconstitution of the DAPK1 tumor suppressor in the format of an immunokinase. After rounds of selections for its most potent cytolytic moiety and optimal ligand part, a DK1KD-SGIII fusion protein containing a constitutive DAPK1 mutant, DK1KD, linked to the scFv SGIII against the B-cell-exclusive endocytic glyco-receptor CD22 was created. Its high purity and large-scale recombinant production provided a stable, selectively binding, and efficiently internalizing construct with preserved robust catalytic activity. DK1KD-SGIII specifically and efficiently killed CD22-positive cells of lymphoma lines and primary CLL samples, sparing healthy donor-or CLL patient-derived non-B cells. The mode of cell death was predominantly PARP-mediated and caspase-dependent conventional apoptosis as well as triggering of an autophagic program. The notoriously high apoptotic threshold of CLL could be overcome by DK1KD-SGIII in vitro also in cases with poor prognostic features, such as therapy resistance. The manufacturing feasibility of the novel CD22-targeting DAPK immunokinase and its selective antileukemic efficiency encourage intensified studies towards specific clinical application. Mol Cancer Ther; 15(5);

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Molecular characterization of neoplastic and normal “sister” lymphoblastoid B-cell lines from chronic lymphocytic leukemia

Cited by 15 publications

References 39 publications

Different expression levels of glycans on leukemic cells—a novel screening method with molecularly imprinted polymers (MIP) targeting sialic acid

Different expression levels of glycans on leukemic cells—a novel screening method with molecularly imprinted polymers (MIP) targeting sialic acid

Cell-Intrinsic Determinants of Ibrutinib-Induced Apoptosis in Chronic Lymphocytic Leukemia

A Novel Recombinant Anti-CD22 Immunokinase Delivers Proapoptotic Activity of Death-Associated Protein Kinase (DAPK) and Mediates Cytotoxicity in Neoplastic B Cells

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