Coexpression of MYC and BCL-2 predicts prognosis in primary gastrointestinal diffuse large B-cell lymphoma

AIM:To investigate whether MYC and BCL-2 coexpression has prognostic significance in primary gastrointestinal diffuse large B-cell lymphoma(PGI-DLBCL)patients,and explore its associations with patients’clinical parameters.METHODS:Fresh and paraffin-embedded tumor tissue samples from 60 PGI-DLBCL patients who had undergone surgery at the Tianjin Medical University Cancer Institute and Hospital from January 2005 to May 2010 were obtained,and 30 lymphoid tissue samples from reactive lymph nodes of age-and sexmatched patients represented control samples.Staging and diagnostic procedures were conducted according to the Lugano staging system.All patients had been treated with three therapeutic modalities:surgery,chemotherapy,or radiotherapy.Expression of MYC and BCL-2 were detected at both protein and m RNA levels by immunohistochemistry and real-time RT-PCR.RESULTS:Positive expression levels of MYC and BCL-2proteins were detected in 35%and 45%of patients,respectively.MYC+/BCL-2+protein was present in30%of patients.MYC and BCL-2 protein levels were correlated with high MYC and BCL-2 m RNA expression,respectively(both P<0.05).We found that advancedstage disease(atⅡE-Ⅳ)was associated with MYC and BCL-2 coexpression levels(P<0.05).In addition,MYC+/BCL-2+patients had more difficulty in achieving complete remission than others(P<0.05).Presenceof MYC protein expression only affected overall survivaland progression-free survival(PFS)when BCL-2 proteinwas coexpressed.The adverse prognostic impact ofMYC+/BCL-2+protein on PFS remained significant(P<0.05)even after adjusting for age,Lugano stage,international prognostic index,and BCL-2 proteinexpression in a multivariable model.CONCLUSION:MYC+/BCL-2+patients have worsechemotherapy response and poorer prognosis thanpatients who only express one of the two proteins,suggesting that assessment of MYC and BCL-2 expressionby immunohistochemistry has clinical significance inpredicting clinical outcomes of PGI-DLBCL patients.

Validation and quality control of hematolymphoid neoplasm immunophenotyping by flow cytometry

cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow for hematopathology.Leukemia/lymphoma immunophenotyping represent an important addition to histomorphology in the diagnosis,classification and monitoring of hematolymphoid neoplasms.The complexity of five-or more parameter analyses and the interpretation of the data rely on standardization and validation of the instrument,the reagent and the procedure.In addition,clinical flow cytometry laboratories in U.S.are required to document proficiency testing,sample preparation,method accuracy,specificity,sensitivity and precision.CLSI and the U.S.-Canadian Consensus Conference have provided recommendations,but each laboratory is responsible for validating its own qualitative and quantitative procedures.This paper introduces the procedures for quality control of all levels of the operation in a clinical flow cytometry laboratory in USA.

The diagnosis and differential diagnosis of granulocytic sarcoma

Objective： To discuss the diagnosis and differential diagnosis of granulocytic sarcoma （GS）. Methods： Six cases were reported in this paper. They were assessed by pathologists. Immunohistochemistry （IHC） stain and routine hematoxylin and eosin （H＆E） stain were applied. Results： All patients involved in different anatomic sites respectively including skin, lymph node, soft tissue, breast, cervix and penis. All cases were previously error diagnoses. Three of them were initially diagnosed as non-Hodgkin lymphoma （NHL）. One case of cervical lymph node lesion was first considered as metastasized carcinoma by clinician. One biopsied skin sample was initially reported as Karposi＇s sarcoma. And one breast case was suspicious of the Iobular carcinoma with the frozen samples without antecedent clinical history information. GS was accompanied with acute myeloid leukemia （AML） in one case and with acute lymphocytic leukemia （ALL） in one case. Histopathologically, blastic, immature and differentiated variants were found in four, one and one, respectively. Immunohistochemistry （IHC） showed that myeloperoxidase （MPO） and lysozyme were both found to be positive in all cases, CD43 was found in 5 of 6 cases. Three of six cases were CD68, CD15 and LCA positive. CD34 and CDl17 were positive in 1/5 and 1/6 cases, respectively. However, CD20 and CD3 were negative in all cases. Conclusion： GS was uncommon and it may be misdiagnosed easily in routine practice. Each area had its own character, but they had the common features too. It can be correctly diagnosed by combination of H＆E stain, IHC stain, peripheral blood and bone marrow. MPO and Lysozyme were necessary for the nature of granulocytes. In addition, CD43, CD68 and CD15 were very helpful.

Manifestations of gastrointestinal plasmablastic lymphoma: A case series with literature review

Plasmablastic lymphoma (PBL) rarely occurs in the gastrointestinal (GI) tract with limited studies reported. We reviewed the clinical histories and pathology of four patients with GI PBL at our institute and similar case reports published in peer-reviewed journals. In our first case, a 40 year-old human immunodeficiency virus positive male presented with a hemorrhoid-like sensation, and was diagnosed with PBL via biopsy of a rectal mass. The second case involves a 65 year-old healthy male with bloody diarrhea who was found to have PBL in a resected sigmoid mass. The third patient was a 41 year-old male with a history of Crohn&#x02019;s disease who presented with abdominal pain, diarrhea, and weight loss. A small intestinal mass (PBL) was removed. The fourth patient was a 65-year-old male who was found PBL after surgical resection of bowel for his florid Crohn&#x02019;s disease. He later developed secondary acute myeloid leukemia. Clinical outcome was very poor in 3 out of 4 patients as reported in the literature. One patient survived chemotherapy followed by autologous transplant. The prototypical clinical presentation and variations of PBL can help create a more comprehensive differential diagnosis for GI tumors and establish an appropriate therapeutic guideline.

癌蛋白NPM-ALK关键磷酸化位点对细胞周期的影响及其机制研究

目的:探讨NPM-ALK蛋白关键磷酸化位点Tyr644和Tyr664对细胞周期的影响及相关的分子机制。方法:建立NPM-ALK和NPM-ALK^(644,664)稳定表达的Jurkat细胞以及瞬时转染的293T细胞,检测各个细胞系的周期分布;应用软琼脂集落形成实验检验野生型NPM-ALK与双点突变型NPM-ALK致瘤能力的差异;利用蛋白质印迹法检测NPM-ALK关键磷酸化位点对下游相关蛋白表达、修饰的调控作用。结果:野生和突变NPM-ALK在瞬时转染的293T细胞和稳定转染的Jurkat细胞中成功表达,转入双点突变型NPM-ALK的Jurkat细胞与转入野生型NPM-ALK的细胞相比呈现出明显的细胞S期阻滞;酪氨酸激酶抑制剂PPP对野生型NPM-ALK转染的Jurkat细胞杀伤效果最强,对双点突变型NPM-ALK转染的细胞的杀伤作用与阴性对照无明显差异;随着NPM-ALK的转入,下游与细胞生长相关分子STAT3、AKT和ERK的磷酸化水平有所升高(P<0.05),双点突变型NPM-ALK转入后,相关蛋白的磷酸化水平与野生型相比显著降低(P<0.05)。结论:癌蛋白NPM-ALK的Tyr644和Tyr664两个关键磷酸化位点的突变会引起细胞S期阻滞和对药物PPP敏感性的降低,其机制可能与NPM-ALK下游的细胞生长相关分子的磷酸化表达变化相关。

仅由t(10;12)(q24;p13)异常所致的难治性急性髓细胞白血病首例报道和文献回顾(英文)

Rearrangements involving chromosome region at 12p13 are common abnormalities in hematological malignancies,including myeloid and lymphoid types. ETV6 gene is usually involved in the 12p13 region. ETV6 rearrangements are more often observed in acute lymphoblastic leukemia than in acute myeloid leukemia (AML),where ETV6 gene deletions are more common than rearrangements. Here,we report an AML case with the recurrent t(10;12)(q24;p13) as the sole abnormality. Fluore-scence in situ hybridization with mapping back to metaphases confirmed that the ETV6 gene splits,and rearranges with a locus at 10q24. In review of the literature,this is the first report of AML case with the novel abnormality as the sole change. Complete laboratory findings from bone marrow examination,flow cytometry analysis,cytogenetic studies,molecular analysis,and clinical features are also described in the report.

Refractory lymphoma treated with chimeric antigen receptor T cells combined with programmed cell death-1 inhibitor:A case report

BACKGROUND Diffuse large B-cell lymphoma(DLBCL)is a common aggressive non-Hodgkin's lymphoma(NHL),accounting for 30%-40%of adult NHL.Primary testicular(PT)lymphoma is an uncommon extranodal disease representing approximately 1%-2%of lymphoma.Approximately 30%–40%of patients are refractory to frontline therapy or relapse after complete remission.Refractory DLBCL responds poorly to other lines of chemotherapy,and experiences short-term survival.CASE SUMMARY We present a 41-year-old male patient who was diagnosed with PT-DLBCL.Further disease progression was observed after multiline chemotherapy.Chimeric antigen receptor T cells(CAR-T)therapy salvaged the patient.Unfortunately,a new mass was observed in the right adrenal area after six months.The patient was administered programmed cell death protein-1(PD-1)inhibitor therapy and maintained progression-free survival at more than 17 mo of follow-up.CONCLUSION Our findings support the potential benefit of CAR-T combined with PD-1 inhibitor therapies in this type of relapsed and refractory PT-DLBCL.

Early postsurgical lethal outcome due to splenic littoral cell angioma:A case report

BACKGROUND Littoral cell angioma(LCA)is a rare benign vascular tumor of the spleen.Given its rarity,standard diagnostic and therapeutic recommendations have yet to be developed for reported cases.Splenectomy is the only method of obtaining a pathological diagnosis and providing treatment to obtain a favorable prognosis.CASE SUMMARY A 33-year-old female presented with abdominal pain for one month.Computed tomography and ultrasound revealed splenomegaly with multiple lesions and two accessory spleens.The patient underwent laparoscopic total splenectomy and accessory splenectomy,and splenic LCA was confirmed by pathology.Four months after surgery,the patient presented with acute liver failure,was readmitted,rapidly progressed to multiple organ dysfunction syndrome and died.CONCLUSION Preoperative diagnosis of LCA is challenging.We systematically reviewed online databases to identify the relevant literature and found a close relationship between malignancy and immunodysregulation.When a patient suffers from both splenic tumors and malignancy or immune-related disease,LCA is possible.Due to potential malignancy,total splenectomy(including accessory spleen)and regular follow-up after surgery are recommended.If LCA is diagnosed after surgery,a comprehensive postoperative examination is needed.

Distinct regulatory mechanism of immunoglobulin gene transcription in epithelial cancer cells

The restriction of immunoglobulin(Ig)expression to B lymphocytes is well established.However,several reports have confirmed that the Ig gene can be expressed in many non-B cancer cells and/or some normal cells.Our aim is to determine whether the Ig gene promoter can be activated in non-B cancer cells and to identify the regulatory mechanism for Ig gene expression.Our results show that the Ig promoter of VH4-59 was activated in several non-B cancer cell lines.Moreover,two novel positive regulatory elements,an enhancer-like element at 2800 to 2610 bp and a copromoter-like element at 2610 to 2300 bp,were identified in two epithelial cancer cell lines,HeLa S3 and HT-29.The octamer element(59-ATGCAAAT-39)located in the Ig promoter,a crucial element for B-cell-derived Ig gene transcription,was also very important for non-B-cell-derived Ig gene transcription.More importantly,we confirmed that octamer-related protein-1(Oct-1),but not Oct-2,was a crucial transcriptional factor for Ig gene transcription due to its ability to bind to the octamer element of the Ig promoter in epithelial cancer cells.These results suggested the presence of a distinct regulatory mechanism for Ig gene expression in non-B cancer cells.

Rearrangement and expression of the immunoglobulin μ-chain gene in human myeloid cells

Immunoglobulin （Ig）, a characteristic marker of B cells, has been reported to be expressed in epithelial cells, with a suggested role in their growth and survival. We have previously reported that IgG heavy chain is expressed in acute myeloid leukemia （AML）, but not in the monocytes or neutrophils from patients with non-hematopoietic neoplasms or healthy controls. In the present study, we assessed IgM heavy chain expression and repertoire in human myeloid cells. We detected VHIzDJHp- rearrangement and expression in 7/7 AML cell lines, 7/14 primary myeloblasts from AML patients, and interestingly, 8/20 monocytes and 3/20 neutrophils from patients with non-hematopoietic neoplasms and healthy individuals. We also found evidence of somatic hypermutation of the variable （V） gene segments in AML-derived IgM gene rearrangements but not in IgM from monocytes or neutrophils from patients with non-hematopoietic neoplasms and healthy individuals. Furthermore, IgM VH^DJH~ gene rearrangements in AML cell lines, primary myeloblasts, and monocytes and neutrophils from patients with non-hematopoietic neoplasms showed a restricted V usage and repertoire, whereas the VH^DJH~ gene rearrangements in monocytes and neutrophils from healthy individuals displayed more diversity. Anti-human IgM inhibited cell proliferation, but did not induce apoptosis in AML cell lines. Our findings suggest that AML-derived IgM might be a novel AML-related molecule that is involved in leukemogenesis and AML progression and might serve as a useful molecular marker for designing targeted therapy and monitoring minimal residual disease.

HDAC inhibitor chidamide synergizes with venetoclax to inhibit the growth of diffuse large B-cell lymphoma via down-regulation of MYC, BCL2, and TP53 expression

Diffuse large B-cell lymphoma(DLBCL) is an aggressive type of non-Hodgkin’s lymphoma. A total of 10%-15% of DLBCL cases are associated with myelocytomatosis viral oncogene homolog(MYC) and/or B-cell lymphoma-2(BCL2) translocation or amplification. BCL2 inhibitors have potent anti-tumor effects in DLBCL;however, resistance can be acquired through up-regulation of alternative anti-apoptotic proteins. The histone deacetylase(HDAC) inhibitor chidamide can induce BIM expression, leading to apoptosis of lymphoma cells with good efficacy in refractory recurrent DLBCL. In this study, the synergistic mechanism of chidamide and venetoclax in DLBCL was determined through in vitro and in vivo models. We found that combination therapy significantly reduced the protein levels of MYC, TP53, and BCL2 in activated apoptotic-related pathways in DLBCL cells by increasing BIM levels and inducing cell apoptosis. Moreover, combination therapy regulated expression of multiple transcriptomes in DLBCL cells, involving apoptosis, cell cycle, phosphorylation, and other biological processes, and significantly inhibited tumor growth in DLBCL-bearing xenograft mice. Taken together, these findings verify the in vivo therapeutic potential of chidamide and venetoclax combination therapy in DLBCL, warranting pre-clinical trials for patients with DLBCL.

Abnormal bone marrow microenvironment: the “harbor” of acute lymphoblastic leukemia cells

Bone marrow(BM)microenvironment regulates and supports the production of blood cells which are necessary to maintain homeostasis.In analogy to normal hematopoiesis,leukemogenesis is originated from leukemic stem cells(LSCs)which gives rise to more differentiated malignant cells.Leukemia cells occupy BM niches and reconstruct them to support leukemogenesis.The abnormal BM niches are the main sanctuary of LSCs where they can evade chemotherapy-induced death and acquire drug resistance.In this review,we focus on the protective effects of BM niche cells on acute lymphoblastic leukemia cells.

Activation of RAS/MAPK pathway confers MCL-1 mediated acquired resistance to BCL-2 inhibitor venetoclax in acute myeloid leukemia

Despite high initial response rates,acute myeloid leukemia(AML)treated with the BCL-2-selective inhibitor venetoclax(VEN)alone or in combinations commonly acquires resistance.We performed gene/protein expression,metabolomic and methylation analyses of isogenic AML cell lines sensitive or resistant to VEN,and identified the activation of RAS/MAPK pathway,leading to increased stability and higher levels of MCL-1 protein,as a major acquired mechanism of VEN resistance.

Critically dysregulated signaling pathways and clinical utility of the pathway biomarkers in lymphoid malignancies

Accumulating evidence confirmed that many dysregulated signaling pathways and aberrant genetic alterations contribute to the oncogenesis and heterogeneity of lymphoid malignancies. Therapeutically targeting dysregulating signaling pathways and their hidden oncogenic biomarkers are becoming available, but did not show desired therapeutic effect in current clinical practice. It is meaningful to further understand the underlying mechanisms of the dysregulated signaling pathways and to address the potential utility of pathway-related biomarkers. To precisely identify the dysregulation of signaling pathways and the 'driver' oncogenic biomarkers, as well as to develop reliable and reproducible risk-stratification based on biomarkers will be challenging. Never-theless, pathway-based targeted therapy will raise the hope to improve the outcomes of the patients with lymphoid malignancies, especially with aggressive types, and the efficient utility of pathway-related biomarkers in diagnosis, prognosis, prediction of lymphoid malignancies may also be able to power precision medicine.

The dichotomy of p53 regulation by noncoding RNAs

The p53 tumor suppressor gene is the most frequently mutated gene in cancer. Significant progress has been made to discern the im- portance of p53 in coordinating cellular responses to DNA damage, oncogene activation, and other stresses. Noncoding RNAs are RNA molecules functioning without being translated into proteins. In this work, we discuss the dichotomy of p53 regulation by noncoding RNAs with four unconventional questions. First, is overexpression of microRNAs responsible for p53 inactivation in the absence of p53 mutation？ Second, are there somatic mutations in the noncoding regions of the p53 gene？ Third, is there a germline mutant in the non- coding regions of the p53 gene that predisposes carriers to cancer？ Fourth, can p53 activation mediated by a noncoding RNA mutation cause cancer？ This work highUghts the prominence of noncoding RNAs in p53 dysregutation and tumorigenesis.

慢性中性粒细胞白血病诊治进展

慢性中性粒细胞白血病(CNL)是一种罕见但可致命的慢性骨髓增殖性肿瘤。其临床特征主要包括以中性粒细胞为主的白细胞升高超过25×109/L,且不伴有病态造血或其他骨髓增殖性肿瘤及生理性中性粒细胞升高。CSF3R T618I突变是CSF3R的激活突变。近期的研究揭示了该突变在CNL发病中的重要作用。目前,CSF3R突变已经被纳入CNL诊断标准。其他突变包括SETBP1及ASXL1,也在疾病发病及进展中发挥着重要作用,且对药物治疗的反应有关。个体化治疗时代,CNL的目标是基于疾病临床特征及分子生物学特征的诊断和治疗。