Contributions
Abstract: EP874
Type: E-Poster Presentation
Session title: Lymphoma Biology & Translational Research
Background
CD47 expresses widely in human tissue and overexpresses in several malignance cells. The interaction between CD47 overexpressing on tumor cells and sSIRPα on macrophages triggers an inhibitory signal cascade that suppresses the phagocytosis function. Berberine is a quaternary amine isoquinoline alkaloid which was traditionally used in intestinal infection and found to have broad anti-tumor activities in several tumors in recent years. However, the function and mechanism of berberine in immunoregulation targeting CD47 is undefined.
Aims
To investigate the functional significance of CD47 in diffuse large B-cell lymphoma (DLBCL) progression and propose an adjuvant agent for DLBCL treatment.
Methods
Paraffin imbedding tissue sections of 55 newly diagnosed DLBCL patients and 40 reactive hyperplasia (RHL) patients were obtained after informed consents. CD47 expression was analyzed in DLBCL tissue sections and DLBCL cell lines. CD47 level was analyzed after berberine treatment, and related regulatory pathway was explored. The immunoregulating function of berberine were proved in co-culture experiment in vitro. A tumor bearing mice model was established to investigate the immunoregulating function of berberine in vivo.
Results
Firstly, we analyzed the CD47 expression in tissue sections form DLBCL and RHL patients. Compared to RHL, CD47 was overexpressed in DLBCL tissues (30/55 vs 11/40, p=0.009, Figure 1A). CD47 positive expression was correlation with higher DLBCL subtype (p=0.007) and Kaplan-Meier analysis showed that patients with positive CD47 expression had a bad OS (Figure 1B). Furthermore, aberrantly increased expression of CD47 was also detected in some DLBCL cell lines (Figure 1C).
Then DLBCL cells were treated with berberine and the results revealed that CD47 was downregulated by berberine at a time-dependent manner (Figure 2A). In the aspect of regulatory signal pathway, CD47 was modulated by berberine at the transcriptional level (Figure 2B). Then, inhibitor of c-myc, a transcriptional factor of CD47, were applied to LY1 cells. The results showed that inhibition of c-myc by 10058-F4 could suppress CD47 expression (Figure 2C, left). The expression level of c-myc was also decreased after berberine treatment (Figure 2C, middle). Moreover, overexpression of c-myc resulted in the increased expression of CD47 and rescued berberine-induced downregulation of CD47 (Figure 2C, right). Engulfment of LY1 cells by macrophage was improved when LY1 cells were pretreated with berberine for 48 hours (Figure 2D). And tumor growth was inhibited by berberine in the tumor-bearing mice model with decreased expression of CD47 and increased infiltrated macrophages (Figure 2E). Moreover, berberine enhanced the efficacy of anti-CD47 antibody and rituximab (Figure 2F).
Conclusion
Our results highlighted that CD47 was a significant feature and played an anti-phagocytosis function in DLBCL progression. We provided evidence for the first time that berberine exerts immunoregulation function in DLBCL via modulating c-myc/CD47 axis and enhance the efficacy of anti-CD47 antibody and rituximab, which provided novel insights into the treatment in DLBCL.
Keyword(s): Diffuse large B cell lymphoma, Immunomodulation, Macrophage
Abstract: EP874
Type: E-Poster Presentation
Session title: Lymphoma Biology & Translational Research
Background
CD47 expresses widely in human tissue and overexpresses in several malignance cells. The interaction between CD47 overexpressing on tumor cells and sSIRPα on macrophages triggers an inhibitory signal cascade that suppresses the phagocytosis function. Berberine is a quaternary amine isoquinoline alkaloid which was traditionally used in intestinal infection and found to have broad anti-tumor activities in several tumors in recent years. However, the function and mechanism of berberine in immunoregulation targeting CD47 is undefined.
Aims
To investigate the functional significance of CD47 in diffuse large B-cell lymphoma (DLBCL) progression and propose an adjuvant agent for DLBCL treatment.
Methods
Paraffin imbedding tissue sections of 55 newly diagnosed DLBCL patients and 40 reactive hyperplasia (RHL) patients were obtained after informed consents. CD47 expression was analyzed in DLBCL tissue sections and DLBCL cell lines. CD47 level was analyzed after berberine treatment, and related regulatory pathway was explored. The immunoregulating function of berberine were proved in co-culture experiment in vitro. A tumor bearing mice model was established to investigate the immunoregulating function of berberine in vivo.
Results
Firstly, we analyzed the CD47 expression in tissue sections form DLBCL and RHL patients. Compared to RHL, CD47 was overexpressed in DLBCL tissues (30/55 vs 11/40, p=0.009, Figure 1A). CD47 positive expression was correlation with higher DLBCL subtype (p=0.007) and Kaplan-Meier analysis showed that patients with positive CD47 expression had a bad OS (Figure 1B). Furthermore, aberrantly increased expression of CD47 was also detected in some DLBCL cell lines (Figure 1C).
Then DLBCL cells were treated with berberine and the results revealed that CD47 was downregulated by berberine at a time-dependent manner (Figure 2A). In the aspect of regulatory signal pathway, CD47 was modulated by berberine at the transcriptional level (Figure 2B). Then, inhibitor of c-myc, a transcriptional factor of CD47, were applied to LY1 cells. The results showed that inhibition of c-myc by 10058-F4 could suppress CD47 expression (Figure 2C, left). The expression level of c-myc was also decreased after berberine treatment (Figure 2C, middle). Moreover, overexpression of c-myc resulted in the increased expression of CD47 and rescued berberine-induced downregulation of CD47 (Figure 2C, right). Engulfment of LY1 cells by macrophage was improved when LY1 cells were pretreated with berberine for 48 hours (Figure 2D). And tumor growth was inhibited by berberine in the tumor-bearing mice model with decreased expression of CD47 and increased infiltrated macrophages (Figure 2E). Moreover, berberine enhanced the efficacy of anti-CD47 antibody and rituximab (Figure 2F).
Conclusion
Our results highlighted that CD47 was a significant feature and played an anti-phagocytosis function in DLBCL progression. We provided evidence for the first time that berberine exerts immunoregulation function in DLBCL via modulating c-myc/CD47 axis and enhance the efficacy of anti-CD47 antibody and rituximab, which provided novel insights into the treatment in DLBCL.
Keyword(s): Diffuse large B cell lymphoma, Immunomodulation, Macrophage