PUBLICATION

2021 Hematological Oncology  
Objective: Diffuse large B cell lymphoma (DLBCL) incidence rates have increased year by year, a part of them have poor clinical outcomes, but the underlying mechanism involved is still unclear. Chemokines have been thought to play an important role in occurrence and development of tumors, but they are poorly studied in DLBCL. CC-Chemokine Ligand 2 (CCL2), the most representative of the CC chemokine family members, through binding to its high affinity receptor, CC chemokine receptor 2 (CCR2),
more » ... be regarded to involve in tumor growth, angiogenesis, epithelial mesenchymal transition, metastasis and immune escape etc. in recent years, but the role and mechanism in DLBCL has not been reported yet. Our preliminary study showed high expression of CCL2 or CCR2 was correlated with clinicopathological characteristics, and an adverse prognostic factor for overall survival (OS) and progression-free survival (PFS) of DLBCL patients. The purpose of this study is to investigate the role of CCL2-CCR2 axis signaling in DLBCL by in vitro experiment. Methods: CCL2 and CCR2 expression were analyzed in human DLBCL cell lines and normal B lymphocytes by Western blot (WB) and qPCR. CCL2 and CCR2 genes were silenced by lentivirus infection. The proliferation, migration, apoptosis and signaling pathway were detected by CCK8, transwell, flow cytometry (FC) and WB, respectively. Results: CCL2 and CCR2 were expressed in all human DLBCL cell lines (SUDHL-2, SUDHL-4, SUDHL-6, OCI-Ly8 and OCI-Ly10). Blockade of CCL2-CCR2 axis signaling with lentivirus infection, CCL2 neutralizing antibody or CCR2 antagonist inhibited tumor cell proliferation, migration and anti-apoptosis ability. The CCL2-CCR2 axis involved in the proliferation and migration of DLBCL cells by activating PI3K/Akt signaling pathway, and induced apoptosis through activation of P38MARK signaling pathway. Conclusions: Our study demonstrates that CCL2/CCR2 axis signaling plays an important role in the development of DLBCL by stimulating cell proliferation, migration and anti-apoptosis. The inhibition of CCL2 or CCR2 may, therefore, be a potential target for anticancer therapy in DLBCL. Background: Acetyl-CoA acetyltransferase 1 (ACAT1) is an enzyme to catalyze reversible formation of acetoacetyl-CoA. Yet, the roles of ACAT1 in malignant progression have been incompletely understood so far. Hence, the aim of this study was to investigate the function and significance of ACAT1 in tumors. Methods: In the present study, clinically annotated tumor patients from multiple cohorts were enrolled with informed consents. Gene expression analysis is completed through the TIMER2 web and GEPIA2 web, based on the combination of the HPA, GTEx, and FANTOM5 datasets. The data of protein expression is acquired from the CPTAC dataset. The statistical significance of survival prognosis is analyzed by log-rank test through the GEPIA2 web. Genetic alteration analysis is accomplished by cBioPortal web. Results: Aberrantly alleviated expression of ACAT1 was detected in multiple tumors at mRNA level compared with normal cells ( Figure 1A ). After including the normal tissue of the GTEx dataset as controls, we further evaluated the expression difference of ACAT1. ACAT1 expression levels were significantly downregulated (tumor versus normal) in tumor cases ( Figure 1B) . Similarly, the outcome of decreased protein level of ACAT1 (tumor versus normal) could be obtained ( Figure 1C ). We divided the cancer cases into highexpression and low-expression groups according to the expression levels of ACAT1 and investigated the correlation of ACAT1 expression with the prognosis of patients with different tumors. Kaplan-Meier curves showed stratified ACAT1 low group were observed with significantly shorter overall survival (OS) and diseasefree survival (DFS) versus the ACAT1 high group in mutiple malignant . wileyonlinelibrary.com/journal/hon tumors ( Figure 1D and 1E). In hematological cancer, ACAT1 high group showed a better overall survival than ACAT1 low group in multiple myeloma and B-cell lymphoma ( Figure 1F ). Then, We observed the genetic alteration status of ACAT1 in different tumor samples of the TCGA cohorts. As shown in Figure 1G , the highest alteration frequency of ACAT1 (>7%) appears for patients with uterine tumors. The "deletion" type of copy number alteration (CNA) was the primary type in melanoma and cervical carcinoma cases, which show an alteration frequency of 2∼3% ( Figure 1G ). The types, sites, and case number of the ACAT1 genetic alteration are further presented in Figure 1H . We found that missense mutation of ACAT1 was the main type of genetic alteration (N=48) ( Figure 1H ). Additionally, we explored the potential association between genetic alteration of ACAT1 and the clinical survival prognosis of cases with different types of cancer. The data of Figure 1I indicated a better overall survival in cases without altered ACAT1 compared with cases with ACAT1 alteration in variety of malignancies. Conclusion: Taken together, the present study was the first pancancer analysis on the role of ACAT1 in tumors. ACAT1 was downregulated and conferred independent prognostic significance, highlighting the effectiveness and potential of ACAT1 as a potential target for future therapeutic strategies. EA - Introduction: Composite lymphoma (CL) is defined as the coexistence of two distinct lymphoma subtypes in a single biopsy, whereas discordant lymphoma (DL) is their occurrence at different anatomical sites. Although CL/DL with two distinct lymphomas are a welldescribed phenomenon, the diagnosis of three lymphomas simultaneously is a much rarer occurrence, with only a few cases reported in the literature. We report a unique case involving two composite lymphomas collectively bearing three distinct diagnostic entities: diffuse large B-cell lymphoma (DLBCL), classic Hodgkin lymphoma (cHL) and follicular lymphoma (FL). Case report: A 48-year-old man presented with cervical lymphadenopathy and a tongue mass. Cervical lymph node biopsy revealed a composite tumour consisting of cHL and DLBCL with a germinal centre B-cell (GCB) phenotype by Hans algorithm. In contrast, the tongue mass biopsy revealed a composite tumour of a morphologically and immunophenotypically similar DLBCL, with FL grade 3B as evidenced by uniform atypical centroblastic cells exhibiting a nodular growth pattern suggestive of neoplastic follicle formation confirmed by intact CD21-positive follicular dendritic cell meshwork. Images highlighting the distinct morphological and immunohistochemical features of all tumour components will be presented. The DLBCL components showed no evidence of MYC rearrangement using break-apart fluorescence in situ hybridization probes. In situ hybridization for Epstein Barr virus RNA was positive in the Hodgkin cells in the cHL component but negative in the other tumours. Although multiplex polymerase chain reaction analysis using BIOMED-2 primers failed to identify clonal rearrangements in IGH, the same clonal IGK rearrangement was detected in all four tumour components, consistent with a shared B-cell clonal origin. The Vk-Kde/intron-Kde reaction identified a single prominent peak at 377 base pair (bp) length amplified by the Vk2f/Vk4/Vk5-Kde primer set, consistent with a clonal B-cell population (Vk-Kde 3, Fig. 1A-C). The same peak was identified in all four tumour components, including the nodal cHL, albeit at a lower amplitude in keeping with the lower tumour cell content of cHL (Fig. 1D). The patient sustained a complete remission following rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone. Conclusion: This case represents an exceedingly rare example of simultaneous composite lymphomas and the concurrent diagnosis of three histologically distinct lymphoma subtypes. It also highlights the limitations of IGH-based clonality testing in germinal centre B-cellderived lymphomas and underscores the importance of adequate tissue sampling to facilitate definitive diagnosis. Given the paucity of data on optimal therapy for these heterogeneous lymphomas, the treatment approach should be individualized, considering all disease components. Introduction: Increasing studies have identified that non-coding RNAs play critical roles in the initiation and progression of tumors via participating in competitive endogenous RNA (ceRNA) networks. However, the roles and functions of the ceRNA network in chronic lymphocytic leukemia (CLL) remain poorly understood. This study aims to explore the molecular mechanism of CLL and provide potential prognostic markers and therapeutic targets through the integrated analysis of the ceRNA network in CLL. Methods: The expression profile of RNAS of CLL patients, CLL cell lines (MEC-1 and EHEB) and healthy group were obtained by the illumina sequencing. R software was used for functional enrichment analysis. The data in the genome microarray map GSE22762 was used for survival analysis. The lncRNA/circRNA-miRNA-mRNA ceRNA networks were visualized by Cytoscape 3.7.2. Four DEmRNAs and the circRNA hsa_circ_0007675/hsa-miR-185-3p/TCF7L1 axis were verified by Quantitative real-time PCR. F I G U R E 1 SUPPLEMENT ABSTRACTS Results: In the present study, 57 differentially expressed-(DE-) mRNAs, 1391 DElncRNAs, 335 DEmiRNAs and 10013 DEcircRNAs were identified comparing CLL patients with healthy donors. Meanwhile, 482 mRNAs, 6085 lncRNAs, 302 miRNAs and 1847 circRNAs were explored differently expressed between CLL cell lines and healthy donors. GO and KEGG analysis results showed that these differentially expressed genes (DEGs) are related to the occurrence and development of tumors. The survival analyses showed that 16 DEGs (INIP, IL3RA, CHD1, NLRP12, IL20RB, B3GALT4, SIT1, ACOT8, AGFGR, PCLAF, C19orf18, SELENOS, F I G U R E 1 388 -SUPPLEMENT ABSTRACTS OR7A17, PCDH7, HNRNPC, PHGDH) were significantly differentially expressed. The ceRNA network were constructed by Cytoscape software. In total, 11 mRNA nodes, 19 miRNA nodes, 105 lncRNA nodes were identified as differentially expressed profiles between CLL patients and control. Meanwhile, a total of 256 DEcircRNAs, 19 DEmiRNAs, and 11 DEmRNAs were employed to construct the ceRNA network between CLL patients and control. We verified four DEGs (TCF7L1, TRIM34, SLC30A10, HOXD4) and the circRNA hsa_circ_0007675/ hsa-miR-185-3p/TCF7L1 axis. Compared with normal people, the expression of these four genes and hsa_circ_0007675 in patient specimens were significantly increased whereas the expression of hsa-miR-185-3p was downregulated (p < 0.05). Conclusion : In this study, we identified the expression profile of RNAs in CLL patients and CLL cell lines. Functional enrichment analysis and survival analysis revealed the potential functions of DEGs. The ceRNA network we established can help to further understand the pathogenesis of CLL and provide potential prognostic markers and novel therapeutic targets. Introduction: Gamma-delta (GD) T-cell lymphoid neoplasms are rare and frequently aggressive tumours whose detection and characterization by flow cytometry (FCM) is challenging in a huge proportion of cases, specially those with leukemic involvement. Sometimes it is difficult to distinguish between clonal/malignant and normal/polyclonal populations. Our aim is to present a case series of GD T-cell entities -focalizing in FCM findings -and review the role of FCM according to the current literature. Methods: Four different and illustrative cases of GD T-cell expansion in peripheral blood (>5% of all lymphocytes) occurred in a singlecenter were reviewed. FCM methodology: eight-colour instrument (FACSCanto TM II) with previous compensation and calibration, EuroFlow panels (screening LST and complete diagnosis T-CLPD. ALOT and T-ALL if necessary), LoQ 0.01%, Infinicyt TM Software 2.0. Normal and clonal/pathological GD T-cells populations were described. Clonally TCR-gamma gene rearrangements detection was performed by multiplex PCR (BIOMED-2). A literature review was performed to describe the characteristics of the most relevant GD T-cell entities.
doi:10.1002/hon.2881 fatcat:fkacb3rv3jexhil4zldos33l2i