Can ELABELA be a novel target in the treatment of chronic lymphocytic leukaemia?

Abstract Background: It has been shown that bcl2, bcl-XL and mcl-1 protein levels are high in chronic lymphocytic leukemia cells, and resultantly, apoptosis does not occur chronic lymphocytic leukemia cells. Apelin and apela (ELABELA/ELA/Toddler) are two peptide ligands for a class A G-protein coupled receptor called apelin receptor. Studies have shown that ELA inhibits apoptosis by inhibiting apoptotic proteins and activating anti-apoptotic proteins. Proteins and genes involved in apoptosis are valuable for targeted cancer therapy. We hypothesized that serum levels may be increased in patients with chronic lymphocytic leukemia based on the antiapoptotic effect of ELA. We compared serum apelin levels of healthy volunteers and patients with chronic lymphocytic leukemia. We aimed to draw attention to a new molecule worthy of research in targeted cancer treatment. Methods: 42 untreated CLL patients and 41 healthy volunteers were included in the study. Serum ELA levels were measured by using enzyme-linked immunosorbent assay kits (Dhanghai Sunred Biological Technology co. Ltd), automated ELISA reader (Thermo Scientific, FİNLAND) and computer program (Scanlt for Multiscan F.C.2.5.1) in accordance with the manufacturer’s instructions. Statistical analysis was done by Statistical Package for Social Sciences for Windows 20 (IBM SPSS Inc., Chicago, IL) ve MedCalc programs. ELA and variables related to CLL were correlated with Spearman correlation anlysis test. ROC analysis and Youden index method were used to determine a cut off point for ELA. All p-values were 2-sided with statistical significance at 0.05 alpha levels. Results: In our study, we found that serum ELA levels were significantly higher in patients with CLL. Conclusions: This study highlights that ELA targeting may be a potential therapeutic option for treating CLL. Keywords: ELABELA; apelinergic system; chronic lymphocytic leukaemia; apoptosis

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Can Apela be a Novel Target in the Treatment of Chronic Lymphocytic Leukaemia?

10.21203/rs.2.11898/v2 ◽

2019 ◽

Author(s):

Didar Yanardag Acik ◽

Mehmet Bankir ◽

Filiz Alkan Baylan ◽

Bilal Aygün

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chronic Lymphocytic Leukaemia ◽

Healthy Volunteers ◽

Lymphocytic Leukaemia ◽

Statistical Significance ◽

Lymphocytic Leukemia ◽

Leukemia Cells ◽

Youden Index ◽

Enzyme Linked Immunosorbent Assay ◽

Apoptotic Proteins

Abstract Background: It has been shown that bcl2, bcl-XL and mcl-1 protein levels are high in chronic lymphocytic leukemia cells, and resultantly, apoptosis does not occur chronic lymphocytic leukemia cells. Apelin and apela (ELABELA/ELA/Toddler) are two peptide ligands for a class A G-protein coupled receptor called apelin receptor. Studies have shown that ELA inhibits apoptosis by inhibiting apoptotic proteins and activating anti-apoptotic proteins. Proteins and genes involved in apoptosis are valuable for targeted cancer therapy. We hypothesized that serum levels may be increased in patients with chronic lymphocytic leukemia based on the antiapoptotic effect of ELA. We compared serum apelin levels of healthy volunteers and patients with chronic lymphocytic leukemia. We aimed to draw attention to a new molecule worthy of research in targeted cancer treatment. Methods: 42 untreated CLL patients and 41 healthy volunteers were included in the study. Serum ELA levels were measured by using enzyme-linked immunosorbent assay kits (Dhanghai Sunred Biological Technology co. Ltd), automated ELISA reader (Thermo Scientific, FİNLAND) and computer program (Scanlt for Multiscan F.C.2.5.1) in accordance with the manufacturer’s instructions. Statistical analysis was done by Statistical Package for Social Sciences for Windows 20 (IBM SPSS Inc., Chicago, IL) ve MedCalc programs. ELA and variables related to CLL were correlated with Spearman correlation anlysis test. ROC analysis and Youden index method were used to determine a cut off point for ELA. All p-values were 2-sided with statistical significance at 0.05 alpha levels. Results: In our study, we found that serum ELA levels were significantly higher in patients with CLL. Conclusions: This study highlights that ELA targeting may be a potential therapeutic option for treating CLL. Keywords: Apela; apelinergic system; chronic lymphocytic leukaemia; apoptosis

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Can Apela be a Novel Target in the Treatment of Chronic Lymphocytic Leukaemia?

10.21203/rs.2.11898/v1 ◽

2019 ◽

Author(s):

Didar Yanardag Acik ◽

Mehmet Bankir ◽

Filiz Alkan Baylan ◽

Bilal Aygün

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chronic Lymphocytic Leukaemia ◽

Healthy Volunteers ◽

Lymphocytic Leukaemia ◽

Statistical Significance ◽

Lymphocytic Leukemia ◽

Leukemia Cells ◽

Youden Index ◽

Enzyme Linked Immunosorbent Assay ◽

Apoptotic Proteins

Abstract Background: It has been shown that bcl2, bcl-XL and mcl-1 protein levels are high in chronic lymphocytic leukemia cells, and resultantly, apoptosis does not occur chronic lymphocytic leukemia cells. Apelin and apela are two peptide ligands for a class A G-protein coupled receptor called apelin receptor. Studies have shown that apella inhibits apoptosis by inhibiting apoptotic proteins and activating anti-apoptotic proteins. Proteins and genes involved in apoptosis are valuable for targeted cancer therapy. We hypothesized that serum levels may be increased in patients with chronic lymphocytic leukemia based on the antiapoptotic effect of apelin. We compared serum apelin levels of healthy volunteers and patients with chronic lymphocytic leukemia. We aimed to draw attention to a new molecule worthy of research in targeted cancer treatment. Methods: 42 untreated CLL patients and 41 healthy volunteers were included in the study. Serum Apela levels were measured by by using enzyme-linked immunosorbent assay (ELISA) kits (Dhanghai Sunred Biological Technology co. Ltd), automated ELISE reader (Thermo Scientific, FİNLAND) and computer program (Scanlt for Multiscan F.C.2.5.1) in accordance with the manufacturer’s instructions. Statistical analysis was made by Statistical Package for Social Sciences (SPSS) for Windows 20 (IBM SPSS Inc., Chicago, IL) ve MedCalc programs. Apela and variables related to CLL were correlated with Spearman correlation anlysis test. ROC analysis and Youden index method were used to determine a cut off point for apela. All p-values were 2-sided with statistical significance at 0.05 alpha levels. Results: In our study, we found that serum apela levels were significantly higher in patients with CLL. Conclusions: This study highlights that apela targeting may be a potential therapeutic option for treating CLL. Keywords: Apela; apelinergic system; chronic lymphocytic leukaemia; apoptosis

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Can ELABELA be a Novel Target in the Treatment of Chronic Lymphocytic Leukaemia?

10.21203/rs.2.11898/v5 ◽

2019 ◽

Author(s):

Didar Yanardag Acik ◽

Mehmet Bankir ◽

Filiz Alkan Baylan ◽

Bilal Aygün

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chronic Lymphocytic Leukaemia ◽

Healthy Volunteers ◽

Lymphocytic Leukaemia ◽

Statistical Significance ◽

Lymphocytic Leukemia ◽

Leukemia Cells ◽

Youden Index ◽

Enzyme Linked Immunosorbent Assay ◽

Apoptotic Proteins

Abstract Background: It has been shown that bcl2, bcl-XL and mcl-1 protein levels are high in chronic lymphocytic leukemia cells, and resultantly, apoptosis does not occur chronic lymphocytic leukemia cells. Apelin and apela (ELABELA/ELA/Toddler) are two peptide ligands for a class A G-protein coupled receptor called apelin receptor. Studies have shown that ELA inhibits apoptosis by inhibiting apoptotic proteins and activating anti-apoptotic proteins. Proteins and genes involved in apoptosis are valuable for targeted cancer therapy. We hypothesized that serum levels may be increased in patients with chronic lymphocytic leukemia based on the antiapoptotic effect of ELA. We compared serum apelin levels of healthy volunteers and patients with chronic lymphocytic leukemia. We aimed to draw attention to a new molecule worthy of research in targeted cancer treatment. Methods: 42 untreated CLL patients and 41 healthy volunteers were included in the study. Serum ELA levels were measured by using enzyme-linked immunosorbent assay kits (Dhanghai Sunred Biological Technology co. Ltd), automated ELISA reader (Thermo Scientific, FİNLAND) and computer program (Scanlt for Multiscan F.C.2.5.1) in accordance with the manufacturer’s instructions. Statistical analysis was done by Statistical Package for Social Sciences for Windows 20 (IBM SPSS Inc., Chicago, IL) ve MedCalc programs. ELA and variables related to CLL were correlated with Spearman correlation anlysis test. ROC analysis and Youden index method were used to determine a cut off point for ELA. All p-values were 2-sided with statistical significance at 0.05 alpha levels. Results: In our study, we found that serum ELA levels were significantly higher in patients with CLL. Conclusions: This study highlights that ELA targeting may be a potential therapeutic option for treating CLL. Keywords: ELABELA; apelinergic system; chronic lymphocytic leukaemia; apoptosis

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Can ELABELA be a Novel Target in the Treatment of Chronic Lymphocytic Leukaemia?

10.21203/rs.2.11898/v3 ◽

2019 ◽

Author(s):

Didar Yanardag Acik ◽

Mehmet Bankir ◽

Filiz Alkan Baylan ◽

Bilal Aygün

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chronic Lymphocytic Leukaemia ◽

Healthy Volunteers ◽

Lymphocytic Leukaemia ◽

Statistical Significance ◽

Lymphocytic Leukemia ◽

Leukemia Cells ◽

Youden Index ◽

Enzyme Linked Immunosorbent Assay ◽

Apoptotic Proteins

Abstract Background: It has been shown that bcl2, bcl-XL and mcl-1 protein levels are high in chronic lymphocytic leukemia cells, and resultantly, apoptosis does not occur chronic lymphocytic leukemia cells. Apelin and apela (ELABELA/ELA/Toddler) are two peptide ligands for a class A G-protein coupled receptor called apelin receptor. Studies have shown that ELA inhibits apoptosis by inhibiting apoptotic proteins and activating anti-apoptotic proteins. Proteins and genes involved in apoptosis are valuable for targeted cancer therapy. We hypothesized that serum levels may be increased in patients with chronic lymphocytic leukemia based on the antiapoptotic effect of ELA. We compared serum apelin levels of healthy volunteers and patients with chronic lymphocytic leukemia. We aimed to draw attention to a new molecule worthy of research in targeted cancer treatment. Methods: 42 untreated CLL patients and 41 healthy volunteers were included in the study. Serum ELA levels were measured by using enzyme-linked immunosorbent assay kits (Dhanghai Sunred Biological Technology co. Ltd), automated ELISA reader (Thermo Scientific, FİNLAND) and computer program (Scanlt for Multiscan F.C.2.5.1) in accordance with the manufacturer’s instructions. Statistical analysis was done by Statistical Package for Social Sciences for Windows 20 (IBM SPSS Inc., Chicago, IL) ve MedCalc programs. ELA and variables related to CLL were correlated with Spearman correlation anlysis test. ROC analysis and Youden index method were used to determine a cut off point for ELA. All p-values were 2-sided with statistical significance at 0.05 alpha levels. Results: In our study, we found that serum ELA levels were significantly higher in patients with CLL. Conclusions: This study highlights that ELA targeting may be a potential therapeutic option for treating CLL. Keywords: ELABELA; apelinergic system; chronic lymphocytic leukaemia; apoptosis

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R-(−)-Gossypol (AT101) Binds to Bcl-2 Family Proteins and Induces Apoptosis in CLL.

Blood ◽

10.1182/blood.v106.11.2978.2978 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2978-2978

Author(s):

Carlos E. Prada ◽

Januario E. Castro ◽

Dayong Zhai ◽

Shinichi Kitada ◽

John C. Reed ◽

...

Keyword(s):

Cell Death ◽

B Cells ◽

Chronic Lymphocytic Leukemia ◽

Programmed Cell Death ◽

Binding Affinity ◽

Lymphocytic Leukemia ◽

Leukemia Cells ◽

Mutational Status ◽

Apoptotic Proteins

Abstract Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of monoclonal B cells in the blood, secondary lymphoid tissues, and marrow. The leukemia cells primarily are arrested in the G0/G1 phase of the cell cycle and appear resistant to programmed cell death. Several anti-apoptotic proteins are over expressed in CLL and this correlates with resistance to treatment, disease progression and overall poor prognosis. Proteins in the Bcl-2 family are central regulators of programmed cell death, and members that inhibit apoptosis, such as Bcl-X(L), Bcl-2, and Mcl-1, are overexpressed in many cancers including CLL and contribute to tumour initiation, progression and resistance to therapy. Mcl-1 is of particular interest because this molecule appears to be regulated by Nurse-like Cells and other stromal cells that promote survival of CLL cells in vitro and very likely also in vivo. These proteins enhance the resistance of CLL cells to spontaneous and/or drug-induced apoptosis primarily by interacting with, and antagonizing the activity of mitochondria membrane pro-apoptotic proteins such as Bax and Bak. The protein-protein interaction of Bcl-2 family members is critical for their activity, and these interactions are governed by binding to the BH3 domain. Racemic gossypol is found in cotton seeds and has been studied as a cytotoxic agent in cancer cell lines and in clinical studies in patients with a large variety of cancers. The antitumor activity of racemic gossypol appears to reside principally in the R-(−)-enantiomer (AT101), with reduced activity observed for the S-(+)-enantiomer. AT101 is an antagonist of the BH3-binding groove of the Bcl-2 family of proteins that can inhibit the interactions of these proteins with pro-apoptotic molecules. We examined whether AT101 could induce apoptosis in Chronic Lymphocytic Leukemia (CLL) and its ability to bind in vitro anti-apoptotic molecules from the Bcl-2 family. Using a Fluorescence Polarization Assay (FPA) we studied the competitive binding affinity of AT101 to Bcl-2 family member proteins. We observed that both, racemic gossypol and AT101 had comparable affinity for Bck-2, Bcl-B, Bfl-1 with EC50=0.6 to 10 μM range. AT101 had a stronger binding affinity to Bcl-X(L) (EC50=0.998 μM vs. 3.084 μM for racemic gossypol), and to Mcl-1 (EC50= 0.52μM vs. 1.07μM for racemic gossypol). CLL cells were incubated with AT101 for 48 hrs at different concentrations. We observed that leukemia cells were induced to undergo apoptosis in a time and dose dependent manner and that this effect was independent of ZAP-70 expression or IgVH gene mutational status (IC50= 2μM). Cells undergoing apoptosis showed PARP-1 cleavage and upregulation of pro-apoptotic molecules such as Bid, p53, as well as downregulation of Mcl-1. These results indicate that AT101 has stronger pan-specific binding affinity for Bcl-2 family proteins than racemic gossypol, in particular to Mcl-1 and Bcl-X(L), and that this compound induces apoptosis in CLL B cells independently of ZAP-70 expression or IgVH gene mutational status. Because of these encouraging results a clinical trial using AT101 in CLL patients with high-risk features is currently open at our institution.

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