Cell Lymphoma Leukemia Research Articles

Cabotegravir, the Long-Acting Integrase Strand Transfer Inhibitor, Potently Inhibits Human T-Cell Lymphotropic Virus Type 1 Transmission in vitro.

Human T-cell lymphotropic virus type 1 (HTLV-1) is a deltaretrovirus most prevalent in southwestern Japan, sub-Saharan Africa, Australia, South America, and the Caribbean. Latest figures approximate 10 million people worldwide to be infected with HTLV-1. This is likely a significant underestimation due to lack of screening in endemic areas and absence of seroconversion symptoms. The two primary diseases associated with HTLV-1 infection are adult T cell leukaemia-lymphoma, a malignant and, sometimes, aggressive cancer; and HTLV-1 associated myelopathy/tropical spastic paraparesis, a debilitating neurological degenerative disease. Unfortunately, despite the poor prognosis, there is currently no effective treatment for HTLV-1 infection. We previously showed that integrase strand transfer inhibitors (INSTIs) clinically used for human immunodeficiency virus type 1 (HIV-1) prophylaxis and treatment are also effective against HTLV-1 transmission in vitro. In 2021 a new INSTI, cabotegravir, was approved by the FDA for HIV-1 treatment. We thus set out to evaluate its efficacy against HTLV-1 infection in vitro. Strand transfer assays performed using recombinant HTLV-1 integrase treated with increasing concentrations of cabotegravir, effectively inhibited strand transfer activity, displaying an IC50 of 77.8 ± 22.4 nM. Furthermore, cabotegravir blocked HTLV-1 transmission in tissue culture; we determined an EC50 of 0.56 ± 0.26 nM, similar to bictegravir. Alu-PCR confirmed the block in integration. Thus, there are four INSTIs and one reverse transcriptase inhibitor approved by the FDA for HIV-1 treatment, that potently block HTLV-1 infection in vitro. This should strongly encourage the establishment of a new standard of HTLV-1 treatment – particularly for pre-exposure prophylaxis and prevention of mother-to-child transmission.

Kiiacylphnols A−H, eight undescribed polycyclic polyprenylated acylphloroglucinols with anticancer activities from Hypericum przewalskii Maxim

Kiiacylphnols A−H, eight previously undescribed polycyclic polyprenylated acylphloroglucinols (PPAPs), along with two known congeners (hyperforcinol F and oxepahyperforin), were obtained from Hypericum przewalskii Maxim. The structures of these metabolites were confirmed by spectroscopic analyses, quantum-chemical 1H and 13C NMR calculations with DP4+ analyses, electronic circular dichroism (ECD) comparisons and calculations. Kiiacylphnols A and B were the first [3.3.1]-type PPAPs with an unusual octahydrooxireno[2,3-i]chromene scaffold bearing a rare 6/6/6/3 ring system. More significantly, kiiacylphnol A and oxepahyperforin displayed cytotoxicity against acute myeloid leukemia and diffuse large B-cell lymphoma cell lines by inducing cell apoptosis.

Clonal Selection and Evolution of HTLV-1-Infected Cells Driven by Genetic and Epigenetic Alteration.

T cells infected with human T-cell leukemia virus type 1 (HTLV-1) acquire various abnormalities during a long latent period and transform into highly malignant adult T-cell leukemia-lymphoma (ATL) cells. This can be described as “clonal evolution”, in which a single clone evolves into ATL cells after overcoming various selective pressures in the body of the infected individuals. Many studies have shown that the genome and epigenome contain a variety of abnormalities, which are reflected in gene expression patterns and define the characteristics of the disease. The latest research findings suggest that epigenomic disorders are thought to begin forming early in infection and evolve into ATL through further changes and accentuation as they progress. Genomic abnormalities profoundly affect clonal dominance and tumor cell characteristics in later events. ATL harbors both genomic and epigenomic abnormalities, and an accurate understanding of these can be expected to provide therapeutic opportunities.

Impact of Human T Cell Lymphotropic Virus Type 1 and 2 Infection on Survival Following Stem Cell Transplantation

Human T cell lymphotropic virus types 1 and 2 (HTLV-1/2) are delta retroviruses. HTLV-1 may lead to complications, including adult T cell leukemia-lymphoma (ATLL) and HTLV-1-associated myelopathy. Immunosuppression may result in progression from an asymptomatic carrier state to ATLL. Data on the safety of stem cell transplantation (SCT) in patients with HTLV-1/2 infection are lacking. The Center for International Blood and Marrow Transplant Research database was queried for patients who tested positive for HTLV infection in the pretransplantation workup and underwent either autologous SCT (autoSCT) or allogeneic SCT (alloSCT). Patients were excluded if they underwent SCT for ATLL. The primary outcome was overall survival (OS) at 3 years and 4 years post-SCT. In those who underwent autoSCT, 54 patients were HTLV-positive and 9836 were HTLV-negative. In those who underwent alloSCT, 105 patients were HTLV-positive and 18,077 were HTLV-negative. No difference in OS was noted between the HTLV-positive and HTLV-negative patients at 3 years post-autoSCT (76% versus 77%; P=.916). Inferior OS (32% versus 46%; P=.017) and nonrelapse mortality (35% versus 27%; P=.030) were observed in HTLV-positive patients at 4 years post-alloSCT. Future work should examine the mechanism by which HTLV-1/2 impact survival in alloSCT recipients.

Research progress in overcoming ibrutinib drug resistance.

Ibrutinib, an oral small-molecule targeted drug, has been the first Bruton tyrosine kinase (BTK) inhibitor in the world to be approved for the market. It works by regulating cell proliferation, apoptosis and migration, and has been proven to exhibit high efficacy and good safety in the treatment of B-cell lymphomas, including chronic lymphocytic leukemia or small lymphocytic lymphoma and mantle cell lymphoma. However, some patients inevitably have drug resistance and disease recurrence, resulting in a poor prognosis. This article serves as a clinical reference by summarizing the related literature on ibrutinib resistance inhibitors.

Prognostic implication of CTLA-4, PD-1, and PD-L1 expression in aggressive adult T-cell leukemia–lymphoma

The prognosis of patients with aggressive adult T cell leukemia-lymphoma (ATLL) is dismal even with intensive chemotherapy. Allogeneic hematopoietic stem cell transplantation (HSCT) is a promising option for patients with aggressive ATLL, but the posttransplant outcome remains unsatisfactory. Hence, to further improve clinical outcomes, novel therapeutic approaches are needed. The clinical significance of immune checkpoint protein expression has not been well-established in aggressive ATLL. This study aims to identify the association between the expression profile of immune checkpoint proteins on ATLL cells and clinical outcomes. This retrospective study cohort included 65 patients with aggressive ATLL diagnosed between 2001 and 2015 at the National Cancer Center Hospital, Tokyo, Japan. Formalin-fixed paraffin-embedded tissue was used to immunohistochemically determine the expression of immune checkpoint proteins and assess the impact of expression profile on the probability of overall survival from diagnosis or HSCT. The current analysis shows that cytotoxic T lymphocyte antigen-4 (CTLA-4), programmed death-1 (PD-1), and programmed death-ligand 1 (PD-L1) expressions were adverse prognostic factors in patients with aggressive ATLL. Experiments that assess the efficacy of immune checkpoint inhibitors are warranted to alleviate the adverse impacts associated with negative immune checkpoints.

CD5-Negative, CD10-Negative Low-Grade B-Cell Lymphoproliferative Disorders of the Spleen.

CD5-negative, CD10-negative low-grade B-cell lymphoproliferative disorders (CD5-CD10-LPD) of the spleen comprise a fascinating group of indolent, neoplastic, mature B-cell proliferations that are essential to accurately identify but can be difficult to diagnose. They comprise the majority of B-cell LPDs primary to the spleen, commonly presenting with splenomegaly and co-involvement of peripheral blood and bone marrow, but with little to no involvement of lymph nodes. Splenic marginal zone lymphoma is one of the prototypical, best studied, and most frequently encountered CD5-CD10-LPD of the spleen and typically involves white pulp. In contrast, hairy cell leukemia, another well-studied CD5-CD10-LPD of the spleen, involves red pulp, as do the two less common entities comprising so-called splenic B-cell lymphoma/leukemia unclassifiable: splenic diffuse red pulp small B-cell lymphoma and hairy cell leukemia variant. Although not always encountered in the spleen, lymphoplasmacytic lymphoma, a B-cell lymphoproliferative disorder consisting of a dual population of both clonal B-cells and plasma cells and the frequent presence of the MYD88 L265P mutation, is another CD5-CD10-LPD that can be seen in the spleen. Distinction of these different entities is possible through careful evaluation of morphologic, immunophenotypic, cytogenetic, and molecular features, as well as peripheral blood and bone marrow specimens. A firm understanding of this group of low-grade B-cell lymphoproliferative disorders is necessary for accurate diagnosis leading to optimal patient management.

Antibacterial and antitumoral properties of 1,2,3-triazolo fused triterpenes and their mechanism of inhibiting the proliferation of HL-60cells.

Antimicrobial resistance and cancer are two important problems affecting human health. Actively developing novel antibiotics and anticancer medicines is a priority. Natural pentacyclic triterpenoids have attracted wide attention due to their significant biological activities. In this study, a series of 1,2,3-triazolo fused triterpenoids (betulin, oleanolic acid and ursolic acid) were functionalized on the A-ring by an in-house developed multi-component triazolization reaction. The compounds were investigated for antitumoral activity in twelve cancer cell lines and were also tested for antibacterial activity against four bacteria. In terms of anticancer effects, compounds 5b-f and 8a-d displayed strong cytotoxic activity in pancreatic adenocarcinoma (Capan-1), chronic myeloid leukemia (Hap-1), acute myeloid leukemia (HL-60), acute lymphoblastic leukemia (Jurkat) and non-Hodgkin lymphoma (Rec-1) cell lines. Among them, compound 5f exhibited the most potent antiproliferative effect on HL-60cells. Further pharmacological research confirmed that compound 5f caused mitochondrial dysfunction and arrested the cell cycle in the G0/G1 phase to induce apoptosis of HL-60cells. In addition, compound 5f also induced autophagy to inhibit the proliferation of HL-60cells. Antibacterial screening revealed that compounds 2a-g and 5a-d showed modest activity against Gram-negative bacteria (Escherichia coli and Salmonella enterica subsp. enterica) with especially compounds 2c and 2d being potent inhibitors of Salmonella enterica subsp. enterica growth. Because of their promising anticancer and antibacterial activity, this series of compounds deserve further study.

The T/Tn-Specific Helix pomatia Lectin Induces Cell Death in Lymphoma Cells Negative for T/Tn Antigens.

Simple SummaryChanges in glycosylation, such as incomplete synthesis and higher density of O-glycans on the cell surface, are frequently observed in cancer cells. Several types of truncated O-glycan structures, e.g., T/Tn antigens, are suspected to disrupt molecular interactions between tumor microenvironment and immune cells, for instance, facilitating cancer immune-escape. Therefore, numerous exogenous lectins targeting aberrant O-glycans are interesting tools for cancer diagnosis, prognosis, and therapy. However, the ability of exolectins to detect subtle alterations in the glycome of tumor cells and to interfere in tumor/healthy cell interactions remains largely unknown. The present article reports for the first time that the Helix pomatia (HPA) lectin, a well-known T/Tn-specific lectin, currently used as a tool in cancer diagnostics, kills Tn-positive leukemia cells and Tn-negative lymphoma cells but does not affect healthy lymphocytes. Thus, HPA could be used to discriminate between tumor and healthy cells, and detect subtle alterations in the glycosylation profile.Morniga G is a T/Tn-specific lectin, inducing cell death in Tn-positive leukemias but not in healthy lymphocytes. Helix pomatia lectin (HPA) is another T/Tn-specific lectin, currently used as tool for cancer diagnostics. The HPA-mediated tumor cell death was evaluated on human leukemia and mouse lymphoma cells, and compared to the effect of Morniga G. Both lectins induced an equivalent percentage of cell death in Tn-positive Jurkat human leukemia. In contrast, EL4 mouse lymphoma resisted Morniga G-mediated cytotoxicity but were killed by HPA at concentrations of 2.5 μg/mL (0.032 nM) and higher. In both malignant cells, HPA-mediated cell death showed features compatible with apoptosis (annexin-externalization, caspase-activation, mitochondrial membrane depolarization, and ROS production). Cytometry analysis indicated that EL4 cells are T/Tn-negative. Because previous results showed a high amount of N-acetylgalactosamine (GalNAc, sugar present in Tn antigen) on EL4 cell surface, this GalNAc could be involved in the formation of truncated O-glycans other than the T/Tn residues. When compared to Morniga G, bioinformatic analysis suggested that HPA benefits from an extended carbohydrate-binding site, better adapted than Morniga G to the accommodation of more complex branched and truncated O-glycans (such as core 2). Finally, HPA killed EL4 cells but not healthy lymphocytes in a mixture of lymphoma cells + lymphocytes, suggesting that HPA selectively triggers tumor cell death.

Chronological genome and single-cell transcriptome integration characterizes the evolutionary process of adult T cell leukemia-lymphoma

Subclonal genetic heterogeneity and their diverse gene expression impose serious problems in understanding the behavior of cancers and contemplating therapeutic strategies. Here we develop and utilize a capture-based sequencing panel, which covers host hotspot genes and the full-length genome of human T-cell leukemia virus type-1 (HTLV-1), to investigate the clonal architecture of adult T-cell leukemia-lymphoma (ATL). For chronologically collected specimens from patients with ATL or pre-onset individuals, we integrate deep DNA sequencing and single-cell RNA sequencing to detect the somatic mutations and virus directly and characterize the transcriptional readouts in respective subclones. Characteristic genomic and transcriptomic patterns are associated with subclonal expansion and switches during the clinical timeline. Multistep mutations in the T-cell receptor (TCR), STAT3, and NOTCH pathways establish clone-specific transcriptomic abnormalities and further accelerate their proliferative potential to develop highly malignant clones, leading to disease onset and progression. Early detection and characterization of newly expanded subclones through the integrative analytical platform will be valuable for the development of an in-depth understanding of this disease.

North American Adult T Cell Leukemia Lymphoma (ATLL) Is Characterized By Distinct, Therapeutically Targetable Mutations in Epigenetic Modifiers

Introduction: We described the mutational spectrum of ATLL in North American patients at ASH 2016 and found that these patients have a heterogenous mutational spectrum (abstract #1758). We also noted some significant differences in comparison to Japanese ATLL cases (Kataoka et al. 2015 PMID: 26437031). In this abstract, we determine the mutational landscape of larger cohort of 27 cases and report on the presence of distinct variants in epigenetic and histone modifying genes.Methods: Twenty-seven North American primary ATLL patient samples (44% acute, 41% lymphomatous and 15% chronic/smoldering) and 7 Japanese cell lines were analyzed by targeted sequencing of 173 recurrently mutated genes in cancer (Genoptix) at a mean sequencing depth of 500X coverage.We then carried out proliferation assays with decitabine on 4 Japanese derived cell lines and 5 primary patient samples and belinostat on 1 cell line and 2 primary patient samples. We also did synergy studies with doxorubicin on 3 sensitive samples. R index was calculated for assessing synergy (Romanelli et al. 1998 PMID: 9523734).Results: We found a high number of epigenetic and histone modifying gene mutations (EP300, TET2, EZH2, MED12, PBRM1, DNMT3A, KMT2A, HIST1H1E, SPEN, IDH1 and ASXL1) in our patients, 15/27 patients (55.5%). Amongst these genes EP300 had the highest frequency of mutations and was present in 22% of patients. (Figure 1) In comparison, the Japanese cohort of 370 cases had 5.7% cases with EP300 mutations. (OR: 4.1; 95% CI: 1.5099 to 10.9492; p = 0.0055)The JAK/STAT pathway had significant number of mutations in the Japanese patients (25% had STAT3 mutations and 5% had JAK3 mutations) whereas in our North American cohort there were no STAT3 mutations and 1 patient had a JAK3 mutation (3.7%). The overall percentage of TP53 mutations was similar in both groups 17.8% in the Japanese and 25.9% in the North American patients. The presence of TP53 mutations was associated with a worse overall survival in our patients (p-value: 0.045) and has been described previously (Magalhaes et al. 2015 PMID: 25573202).EP300 is a histone acetyltransferase that acetylates histones and leads to activation of gene transcription. Inactivating mutations in EP300 can lead to aberrant gene silencing and can be associated with increased DNA methylation. A high prevalence of EP300 mutations prompted us to test the sensitivity of ATLL cell lines and primary samples to DNA methyltransferase inhibitor (Decitabine) and histone deacetylase inhibitor (Belinostat). Among the 4 cell lines tested, only the ATL43Tb(-) cell line with EP300 mutation was sensitive to the growth inhibitory effects of decitabine (IC50 < 1 uM) whereas the IC50 was not reached in resistant samples (three Japanese derived cell lines, ED40515(-) and ED41214(-) and Su9T01(-)) (Figure 2A). Since the cell lines were derived from Japanese ATLL, next, we established long term cultures of primary ATLL cells from North American cases and used them for functional studies. For primary samples, we observed that all samples (n=5) were sensitive to decitabine with IC50 < 1 uM. Furthermore, the three samples (EP300 mutant) were also sensitive to HDAC inhibitor Belinostat with an IC50 of upto 0.5 uM in all samples. Next, we tested the efficacy of combining decitabine with chemotherapy. The combination of doxorubicin and decitabine showed synergy with an R index >1 in three EP300 mutant samples tested Pt-5a, Pt-7a and ATL43Tb(-).Conclusions: Mutational analysis of the largest cohort of North American ATLL samples shows a distinct variant landscape with a high prevalence of mutations affecting epigenetic modifiers. EP300 is a histone acetyltransferase that is mutated in 22% of samples and is significantly higher in the North American cohort when compared to the Japanese cases.We also determined that majority of primary ATLL samples and EP300 mutant cell lines tested are sensitive to decitabine. The combination with doxorubicin shows synergy. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Deletion 7q Is Associated with KLF2 and NOTCH2 Mutations and Is Strongly Correlated with Splenic Marginal Zone Lymphoma but Also Found in Lymphoplasmacytic Lymphoma and Hairy Cell Leukemia Variant

Introduction: Deletion 7q (del(7q)) isa common cytogenetic aberration in lymphoma and has been associated with splenic marginal zone lymphoma (SMZL). Del(7q) has been proposed to distinguish SMZL from other forms of MZL, i.e. MALT lymphoma and nodal MZL. However, the value of del(7q) in the differential diagnosis of other lymphoid malignancies has not been elucidated. Furthermore, the overall increasing diagnostic and prognostic importance of somatic mutations in hematologic malignancies prompted us to analyze the mutational landscape in lymphomas with del(7q).Aims: To understand the role of del(7q) in lymphoma diagnosis, to define the minimal deleted region in del(7q) bearing lymphomas and to analyze co-occurring cytogenetic aberrations and co-mutations.Patients and Methods: From 2005 to 2017 a total of 5251 patients were diagnosed with lymphoma at our institution by flow cytometry, chromosome banding analysis (CBA) and FISH (2547 (48%) in bone marrow, 2704 (52%) in peripheral blood). Subject of the present study were 3917 patients who were selected based on an aberrant karyotype by (CBA) in order to exclude inclusion of false negative CBA due to limited lymphoma infiltration. To evaluate del(7q) as biological relevant primary hit, del(7q) had to be part of the major clone without a complex karyotype (≥3 abnormalities). 109/3917 patients met this criterion. CD5 negative lymphomas were diagnosed by flow cytometry according to the respective suggestive phenotype as SMZL or lymphoplasmacytic lymphoma (LPL). For 37/109 patients (34%) that had ≥ 20% of lymphoma infiltration by flow cytometry the diagnostic sample was available for array comparative genomic hybridization (aCGH). For 44/109 patients (40%) that had ≥10% infiltration by flow cytometry the diagnostic sample was available for sequencing of 63 genes frequently mutated in hematologic malignancies.Results: We found 109/3917 patients with a del(7q) by CBA (3%). Lymphoma diagnosis with del(7q) included the following eight entities as defined by flow cytometry at decreasing frequency: SMZL (59%), LPL (21%), B-cell prolymphocytic leukemia (6%), hairy cell leukemia variant (HCLv, 6%), chronic lymphocytic leukemia (CLL, 4%), mantle cell lymphoma (2%), aggressive lymphoma (2%), and follicular lymphoma (0.9%). The presence of del(7q) was significantly associated with SMZL, LPL and HCLv (each p<0.0001). Del(7q) was negatively correlated with CLL (p<0.0001). Recurrent cytogenetic aberrations that co-occurred with del(7q) include del(11q), trisomy 3 and dup(1q) in 9%, 3% and 2%, respectively.We analyzed 37/109 patients with aCGH and defined a minimal deleted region of 1.9 MB (Chr7; 128.973.948-130.908.181). Deleted genes in this region include KLF14 (kruppel-like factor 14) and a microRNA (MIR) cluster associated with tumor suppression (MIR182-96-183, MIR 29A, MIR96). The deletion was not entity specific, but 2/5 CLL samples showed a deletion outside the common region of the other lymphomas (7q35-q36.3 and 7q11.21-q21.2).In 44/109 patients the diagnostic sample was analyzed by gene panel sequencing (63 genes) to identify the mutational landscape of del(7q) lymphoma. We identified a total of 53 mutations in 19 different genes in 29/44 patients (66%). The top 10 mutated genes were: KLF2 (27%), NOTCH2 (23%), CXCR4 (9%), MYD88 (9%), NOTCH1 (7%), BIRC3 (7%), MAP2K1 (5%), NRAS (5%), TET2 (5%) and TP53 (5%). Co-occurring mutations in KLF2 and NOTCH2 were observed in 4/44 patients (9%).Somatic mutations in NOTCH2 (10/10) and/or KLF2 (12/12) were exclusively observed in SMZL, and MYD88 (4/4) mutations exclusively in LPL as described in the literature (Fig.). The presence of del(7q) and a mutation in KLF2 and/or NOTCH2 was highly predictive for the diagnosis of SMZL (p<0.0001). Moreover in the presence of del(7q) 18/44 analyzed patients (41%) had NOTCH2 or KLF2 mutations compared to 8% in the control cohort (p<0.0001).Conclusion: Our analysis shows that del(7q) is not restricted to SMZL and is in fact associated also with LPL and HCLv. There is a small common deleted region spanning 1.9 MB at 7q31.32q32.3 which is not entity specific and includes a MIR cluster associated with tumor suppression. Mutations in KLF2 and NOTCH2 were found to be associated with del(7q), which raises the possibility of a common mechanism that remains to be elucidated. Taken together del(7q) combined with somatic mutations in KLF2 and/or NOTCH2 is highly specific for SMZL. [Display omitted] DisclosuresHöllein:MLL Munich Leukemia Laboratory: Employment. Stengel:MLL Munich Leukemia Laboratory: Employment. Meggendorfer:MLL Munich Leukemia Laboratory: Employment. Fasan:MLL Munich Leukemia Laboratory: Employment. Kern:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership.

Ethiopian Medicinal Plants Traditionally Used for the Treatment of Cancer; Part 3: Selective Cytotoxic Activity of 22 Plants against Human Cancer Cell Lines.

Medicinal plants have been traditionally used to treat cancer in Ethiopia. However, very few studies have reported the in vitro anticancer activities of medicinal plants that are collected from different agro-ecological zones of Ethiopia. Hence, the main aim of this study was to screen the cytotoxic activities of 80% methanol extracts of 22 plants against human peripheral blood mononuclear cells (PBMCs), as well as human breast (MCF-7), lung (A427), bladder (RT-4), and cervical (SiSo) cancer cell lines. Active extracts were further screened against human large cell lung carcinoma (LCLC-103H), pancreatic cancer (DAN-G), ovarian cancer (A2780), and squamous cell carcinoma of the esophagus (KYSE-70) by using the crystal violet cell proliferation assay, while the vitality of the acute myeloid leukemia (HL-60) and histiocytic lymphoma (U-937) cell lines was monitored in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) microtiter assay. Euphorbia schimperiana, Acokanthera schimperi, Kniphofia foliosa, and Kalanchoe petitiana exhibited potent antiproliferative activity against A427, RT-4, MCF-7, and SiSo cell lines, with IC50 values ranging from 1.85 ± 0.44 to 17.8 ± 2.31 µg/mL. Furthermore, these four extracts also showed potent antiproliferative activities against LCLC-103H, DAN-G, A2780, KYSE-70, HL-60, and U-937 cell lines, with IC50 values ranging from 0.086 to 27.06 ± 10.8 µg/mL. Hence, further studies focusing on bio-assay-guided isolation and structural elucidation of active cytotoxic compounds from these plants are warranted.

Establishment and Usage of Leukemia-Lymphoma Cell Lines--Review

Cancer cell lines are an indispensable tool in the cancer research. Since the first human cell line, HeLa was established in the 1950s, thousands of cancer cell lines have been established, including 637 characterized leukemia-lymphoma cell lines. The probability to successfully establish cancer cell lines is a low by traditional methods, and the addition of regulatory factors is often required. However, a novel "conditional reprogramming" technology can improve this situction. The establishment and description of a new cell line should be consistent with international guidelines. Cancer cell lines are mainly used in the research of tumor pathogenesis and drug development. Scientists have developed many kinds of cell line panels which can be used for the high-throughput screening of anticancer drugs. Mycoplasma contamination and/or cross-contamination from other cells should be avoided during the use of cell lines. The establishment of a cell model passport database can prevent those misidentifications. In this review, the types, establishment and usage of leukemia-lymphoma cell lines as well as points of attention when using them are summarized briefly.

Diffuse large B-cell lymphoma with leukemic involvement.

e19552 Background: Leukemic involvement (LI) in diffuse large B cell lymphoma (L-DLBCL) is very rare and has been sparsely reported. We report our experience with this entity in a large academic multi-center setting. Methods: Medical records of patients with DLBCL who received care at Mayo Clinic between 1/2003 and 6/2020 were reviewed. DLBCL patients with LI were identified. LI was defined as increased white blood cell counts and the presence of clonal B cells on peripheral blood flow-cytometry. Kaplan-Meier method was used for survival analysis. Results: Twenty patients with L-DLBCL were identified with a median follow-up of 32.5 months (CI95%, 32.5-NR). Median age at initial diagnosis was 62 (45-80) years. 60% (12/20) were male; 90% (18/20) were Caucasian. Pathologically, 90% (18/20) had DLBCL and 10% (2/20) had high-grade B cell lymphoma (HGBCL) with intermediate features between DLBCL and Burkitt lymphoma. By Hans criteria, 58% (11/19) had germinal center B-cell DLBCL (GCB-DLBCL) and 42% (8/19) had non-germinal center B-cell DLBCL (non-GCB-DLBCL). 40% (8/20) had transformed DLBCL (t-DLBCL); 36% (5/14) had double-hit lymphoma (DHL) by FISH analysis. LI was present in 55% (11/20) at initial diagnosis and 45% (9/20) at relapse. Median WBC was 39.5/ul (range, 4.3-121) with median absolute lymphocyte count of 25 k/ul (range, 0.7-117). Immunophenotypically, the leukemic lymphoma cells expressed CD19, CD20, and CD79a. Bone marrow involvement and pancytopenia were documented in all patients with a median bone marrow cellularity of 80%. Other extranodal sites of involvement with LI included spleen (65%;13/20), liver (20%;4/20), breast and soft tissue (20%;4/20), bladder or kidneys (10%;2/20), skeleton (10%;2/20), and myocardium (5%;1/20). 65% (13/20) had B-symptoms. All patients had LDH elevation (UNL 222 U/L) with a median of 2125 U/L (range, 308-10,760). 45% (5/11) of patients with LI at initial diagnosis had CNS involvement on relapse/progression. All patients with LI at initial diagnosis received anthracycline-based chemoimmunotherapy with or without CNS prophylaxis. Patients with LI at relapse had had a median of 2 prior treatments (range, 1-5) before LI. Median overall survival (OS) for the whole group was 9 months (CI 95%; 5.8-11.8). There were no long-term survivors. Median progression free survival after LI was 4.7 months (CI95%; 0.8-7.6) in the newly diagnosed group and 3 months (CI95%; 0.9-20) in the relapsed group. 90% (18/20) died due to their progressive disease. Cell of origin, DHL status, or newly diagnosed vs. relapsed status did not have a significant impact on OS in patients with L-DLBCL. Conclusions: Leukemic involvement at any time during the course of DLBCL is associated with poor prognosis. It also appears to be a major risk factor for CNS relapse. It is most frequently associated with DHL and t-DLBCL. Novel therapeutic approaches at the time of initial therapy need to be developed for L-DLBCL.

Épidémiologie des syndromes lymphoprolifératifs chroniques B

AbstractIntroduction. There is an increasing incidence of cancer worldwide. Geographic and ethnic disparities are identified, with a higher incidence and poorer survival among African Americans compared to Whites.Goal. Presentation of incidence and survival data for the seven major B-lymphoproliferative syndromes (SLP-B): multiple myeloma (MM), chronic lymphocytic leukemia (CLL), follicular lymphoma (FL), marginal zone lymphoma (LZM), lymphoma lymphoplasmacytic (LPL), mantle cell lymphoma (LCM) and hairy cell leukemia (LT).Results. It is the elderly who is most frequently affected. In 2018, in France the number of incident cases was estimated at 17,163 cases, including 5,442 patients with MM, 4,674 with CLL, 3,066 with LF, 2,790 with LZM, 1.317 with LPL, 887 with LCL and 304 with LT. Incidence rates are increasing over recent periods for MM, LF, LZM but on the other hand decreases for CLL and LPL while the incidence of LT remains stable. Hematological malignancies with an intermediate prognosis are MM and MCL, with a 5-year standardized net survival of 60% and 63%, respectively. The prognosis is good with survival> 90% (95%) for LT, close to 90% for CLL (89%), LF (86%) and LZM (88%) and close to 80% (82%) for the LZM. If LZM is associated with hepatitis C, the increase in incidences is more linked to the population structure and its aging but also to changes in practice with easier access to protein electrophoresis and flow cytometry.Conclusion. Epidemiological studies are important for better understanding the diseases, analyzing the temporal variations observed and assessing the therapeutic progress made.

Direct single-molecule imaging for diagnostic and blood screening assays

Every year, over 100 million units of donated blood undergo mandatory screening for HIV, hepatitis B, hepatitis C, and syphilis worldwide. Often, donated blood is also screened for human T cell leukemia-lymphoma virus, Chagas, dengue, Babesia, cytomegalovirus, malaria, and other infections. Several billion diagnostic tests are performed annually around the world to measure more than 400 biomarkers for cardiac, cancer, infectious, and other diseases. Considering such volumes, every improvement in assay performance and/or throughput has a major impact. Here, we show that medically relevant assay sensitivities and specificities can be fundamentally improved by direct single-molecule imaging using regular epifluorescence microscopes. In current microparticle-based assays, an ensemble of bound signal-generating molecules is measured as a whole. By contrast, we acquire intensity profiles to identify and then count individual fluorescent complexes bound to targets on antibody-coated microparticles. This increases the signal-to-noise ratio and provides better discrimination over nonspecific effects. It brings the detection sensitivity down to the attomolar (10-18 M) for model assay systems and to the low femtomolar (10-16 M) for measuring analyte in human plasma. Transitioning from counting single-molecule peaks to averaging pixel intensities at higher analyte concentrations enables a continuous linear response from 10-18 to 10-5 M. Additionally, our assays are insensitive to microparticle number and volume variations during the binding reaction, eliminating the main source of uncertainties in standard assays. Altogether, these features allow for increased assay sensitivity, wide linear detection ranges, shorter incubation times, simpler assay protocols, and minimal reagent consumption.

Polyphyllin D Shows Anticancer Effect through a Selective Inhibition of Src Homology Region 2-Containing Protein Tyrosine Phosphatase-2 (SHP2).

Natural products have continued to offer tremendous opportunities for drug development, as they have long been used in traditional medicinal systems. SHP2 has served as an anticancer target. To identify novel SHP2 inhibitors with potential anticancer activity, we screened a library containing 658 natural products. Polyphyllin D was found to selectively inhibit SHP2 over SHP1, whereas two other identified compounds (echinocystic acid and oleanolic acid) demonstrated dual SHP1 and SHP2 inhibition. In a cell-based assay, polyphyllin D exhibited cytotoxicity in Jurkat cells, an acute lymphoma leukemia cell line, whereas the other two compounds were ineffective. Polyphyllin D also decreased the level of phosphorylated extracellular signal-regulated kinase (p-ERK), a proliferation marker in Jurkat cells. Furthermore, knockdown of protein tyrosine phosphatase (PTP)N6 (SHP1) or PTPN11 (SHP2) decreased p-ERK levels. However, concurrent knockdown of PTPN6 and PTPN11 in Jurkat cells recovered p-ERK levels. These results demonstrated that polyphyllin D has potential anticancer activity, which can be attributed to its selective inhibition of SHP2 over SHP1.

The Paradoxical Roles of Orphan Nuclear Receptor 4A (NR4A) in Cancer

The three-orphan nuclear receptor 4A genes are induced by diverse stressors and stimuli, and there is increasing evidence that NR4A1 (Nur77), NR4A2 (Nurr1), and NR4A3 (Nor1) play an important role in maintaining cellular homeostasis and in pathophysiology. In blood-derived tumors (leukemias and lymphomas), NR4A expression is low and NR4A1-/-/NR4A3-/- double knockout mice rapidly develop acute myelocytic leukemia, suggesting that these receptors exhibit tumor suppressor activity. Treatment of leukemia and most lymphoma cells with drugs that induce expression of NR4A1and NR4A3 enhances apoptosis, and this represents a potential clinical application for treating this disease. In contrast, most solid tumor-derived cell lines express high levels of NR4A1 and NR4A2, and both receptors exhibit pro-oncogenic activities in solid tumors, whereas NR4A3 exhibits tumor-specific activities. Initial studies with retinoids and apoptosis-inducing agents demonstrated that their cytotoxic activity is NR4A1 dependent and involved drug-induced nuclear export of NR4A1 and formation of a mitochondrial proapoptotic NR4A1-bcl-2 complex. Drug-induced nuclear export of NR4A1 has been reported for many agents/biologics and involves interactions with multiple mitochondrial and extramitochondrial factors to induce apoptosis. Synthetic ligands for NR4A1, NR4A2, and NR4A3 have been identified, and among these compounds, bis-indole derived (CDIM) NR4A1 ligands primarily act on nuclear NR4A1 to inhibit NR4A1-regulated pro-oncogenic pathways/genes and similar results have been observed for CDIMs that bind NR4A2. Based on results of laboratory animal studies development of NR4A inducers (blood-derived cancers) and NR4A1/NR4A2 antagonists (solid tumors) may be promising for cancer therapy and also for enhancing immune surveillance.

Progress in the treatment of NK-cell lymphoma/leukemia

Natural killer (NK)/T cell lymphoma includes two major subtypes of disease, specifically extranodal NK/T cell lymphoma, nasal type (ENKL) and aggressive NK cell leukemia (ANKL). Both are strongly associated with Epstein-Barr virus and are prevalent in East Asia and Latin America. Except for that of limited-stage ENKL, the prognosis of both diseases was poor in the previous decade. The advent of non-anthracycline-based chemoradiotherapy has contributed to an improvement in ENKL prognosis, but there is still room for further treatment progress. Recently, the high efficacy of PD-1 antibody was reported in relapsed or refractory ENKL patients. This was later supported by the finding that PD-L1/PD-L2 genetic alterations are frequently observed in ENKL and ANKL patients. Due to the rarity of the disease, a standard treatment for ANKL remains to be established. Currently, allogeneic stem cell transplantation is the only curative treatment, and this is even applicable to chemo-resistant ANKL patients. In this review, we focus on recent treatment approaches for NK/T cell lymphomas including novel agents.