Comparative analysis of testis transcriptomes associated with male infertility in triploid cyprinid fish

2019 ◽  
Vol 31 (2) ◽  
pp. 248 ◽  
Author(s):  
Wuhui Li ◽  
Hui Tan ◽  
Junmei Liu ◽  
Jie Hu ◽  
Jialin Cui ◽  
...  
Keyword(s):  
Male Infertility ◽  
B Cell ◽  
Ring Finger ◽  
B Cell Lymphoma ◽  
Diploid Parent ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Caspase 7 ◽  
Ring Finger Proteins ◽  
Ubiquitin Conjugating Enzymes

Spermatogenesis involves a series of cellular transformations and thousands of regulated genes. Previously, we showed that the triploid fish (3nBY) cannot produce mature spermatozoa. In the present study, evaluation of the testis microstructure revealed that germ cells of 3nBY could develop into round spermatids, but then degenerated, resulting in male infertility. In this study we comparatively analysed the testis transcriptomes from 3nBY and its diploid parent YB and identified a series of differentially expressed genes (DEGs) that were enriched in the Wnt signalling pathway and the apoptotic and ubiquitin-mediated proteolysis processes in 3nBY. Gene ontology functional analyses revealed that some DEGs in 3nBY were directly associated with the process of gamete generation, development and sperm flagellum assembly. In addition, the expression of a number of genes related to meiosis (Inhibitor Of DNA Binding 2 (ID2), Ovo Like Transcriptional Repressor 1 (OVOL1)), mitochondria (ATP1b (ATPase Na+/K+ Transporting Subunit Beta 1), ATP2a (ATPase, Ca++ Transporting, Cardiac Muscle, Slow Twitch 2), ATP5a (ATP Synthase F1 Subunit Alpha), Mitochondrially Encoded Cytochrome C Oxidase I (COX1), NADH Dehydrogenase Subunit 4 (ND4)) and chromatin structure (Histone 1 (H1), Histone 2a (H2A), Histone 2b (H2B), Histone 3 (H3), Histone 4 (H4)) was lower in the testes of 3nBY, whereas the expression of genes encoding ubiquitin (Ubiquitin Conjugating Enzymes (UBEs), Ring Finger Proteins (RNFs)) and apoptosis (CASPs (Caspase 3, Caspase 7,Caspase 8), BCLs (B-Cell Lymphoma 3, B-Cell CLL/Lymphoma 2, B Cell CLL/Lymphoma 10)) proteins involved in spermatid degeneration was higher. These data suggest that the disrupted expression of genes associated with spermatogenesis and the increased expression of mitochondrial ubiquitin, which initiates cell apoptosis, may result in spermatid degeneration in male 3nBY. This study provides information regarding the potential molecular regulatory mechanisms underlying male infertility in polyploid fish.

IL-4 Affects Proliferation, Chemosensitivity-and Rituximab Sensitivity of Germinal Center B-Cell like (GCB) and Activated B-Cell like (ABC) Diffuse Large B-Cell Lymphoma Differently.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 242-242 ◽  
Author(s):  
Hovav Nechushtan ◽  
Joseph D. Rosenblatt ◽  
Izidore S. Lossos
Keyword(s):  
Cell Death ◽  
B Cell ◽  
Cell Lines ◽  
Germinal Center ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Cell Lysis ◽  
Large B Cell Lymphoma ◽  
Expression Of Genes ◽  
Large B Cell

Abstract Diffuse Large B-cell Lymphoma (DLBCL) represent a diverse group of lymphoid neoplasms with heterogeneous clinical, histological, immunophenotypic, cytogenetic and molecular genetic features. Approximately 50% of DLBCL patients are not cured by the standard combination chemotherapy regimens. DLBCL can be subclassified into GCB-like DLBCL which are characterized by expression of genes normally expressed in germinal center B cells, and having a significantly better overall survival (OS) than the ABC-like DLBCL, which are characterized by expression of genes induced during in vitro activation of normal B cells. At least two markers of the GCB-phenotype - BCL6 and HGAL - are IL-4 target genes, increased expression of which independently predicts better OS. These observations suggest that endogenous or exogenously administered IL-4 may influence behavior of DLBCL. IL-4 mRNA was detected at low levels in 5 of 7 GCB-like and in all 4 ABC-like DLBCL tumor specimens. Two of 7 GCB-like tumors showed high expression levels of IL-4 as determined by real-time RT-PCR. Examination of the effects of IL-4 on proliferation of GCB-like (SUDHL6, SUDHL4 and OCILY19) and ABC-like (OCILY10 and OCILY3) DLBCL cell lines showed that IL-4 mildly increased DNA synthesis, as assessed by thymidine incorporation, in all the GCB-like DLBCL. Conversely, IL-4 markedly decreased proliferation in the ABC-like DLBCL cell lines by inducing G1 arrest. IL-4 also differently affected the sensitivity of GCB-like and ABC-like DLBCL to doxorubicin. IL-4 reduced doxorubicin-induced cell death of ABC-like cell lines (20–50% reduction) while it markedly increased the killing of the GCB-like cells (40–80% induction). IL-4 also prevented serum starvation-induced cell death of the ABC-like DLBCL, but it increased cell death of the GCB-like DLBCL cell lines. Recently, Rituximab was shown to improve survival of DLBCL patients when added to the CHOP regimen. The precise mechanisms of its action are unknown; however present data suggest that it may affect lymphoma cells either by activation of complement lysis or by mediating ADCC. IL-4 reduced the complement mediated Rituximab cell lysis of the ABC-like cell lines, while it increased the complement mediated Rituximab cell lysis of the GCB-like DLBCL cell lines. Expression levels of surface markers that modulate complement cell lysis (CD46, CD55 and CD59) were not affected by IL-4 exposure. In contrast, IL-4 did not affect killing of GCB-like and ABC-like cells by ADCC. These observations suggest that DLBCL subtypes may respond differently to the in vivo cytokine milieu of the tumor. Different responsiveness to IL-4 may modulate tumor sensitivity to the current therapeutic modalities and can potentially be explored to augment response to chemotherapy and Rituximab.

scholarly journals Ubiquitin Ligase Activities of Bombyx mori Nucleopolyhedrovirus RING Finger Proteins

2003 ◽  
Vol 77 (2) ◽  
pp. 923-930 ◽  
Author(s):  
Noriko Imai ◽  
Noriyuki Matsuda ◽  
Keiji Tanaka ◽  
Akihiko Nakano ◽  
Shogo Matsumoto ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Ubiquitin Ligase ◽  
Mutational Analysis ◽  
Ring Finger ◽  
Zinc Ion ◽  
Ubiquitin Ligase E3 ◽  
Reticulocyte Lysates ◽  
Ring Finger Proteins ◽  
Ubiquitin Conjugating Enzymes

ABSTRACT The genome of Bombyx mori nucleopolyhedrovirus (BmNPV) is predicted to contain six RING finger proteins: IAP1, ORF35, IAP2, CG30, IE2, and PE38. Several other members of the RING finger family have recently been shown to have the ubiquitin-ligase (E3) activity. We thus examined whether BmNPV RING finger proteins have the E3 activity. In vitro ubiquitination assay with the rabbit reticulocyte lysates and BmNPV RING finger proteins fused with maltose-binding protein (MBP) showed that four of them (IAP2, IE2, PE38, and CG30) were polyubiquitinated in the presence of zinc ion. Furthermore, MBP-IAP2, MBP-IE2, and MBP-PE38 were able to reconstitute ubiquitination activity in cooperation with the Ubc4/5 subfamily of ubiquitin-conjugating enzymes. Mutational analysis also showed that ubiquitination activity of MBP-IAP2, MBP-IE2, and MBP-PE38 were dependent on their RING finger motif. Therefore, these results suggest that IAP2, IE2, and PE38 may function as E3 enzymes during BmNPV infection.

scholarly journals Differential Expression of TCF3 Target Genes Defines Subclasses of Diffuse Large B-Cell Lymphoma with Striking Differences in Clinical Outcome Following R-CHOP Therapy

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3037-3037 ◽  
Author(s):  
Kathrin Tyryshkin ◽  
Yi D Li ◽  
David Good ◽  
Lois E. Shepherd ◽  
Tara Baetz ◽  
...  
Keyword(s):  
B Cell ◽  
Differential Expression ◽  
Target Genes ◽  
De Novo ◽  
B Cell Lymphoma ◽  
Large B Cell Lymphoma ◽  
Expression Of Genes ◽  
Pre Treatment ◽  
Differential Expression Of Genes ◽  
Chop Therapy

Abstract Background: Diffuse large B-cell lymphoma (DLBCL) is clinically and biologically heterogeneous. Whereas most patients are cured following conventional therapy with rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP), a substantial minority are refractory or relapse. Identifying patients at high risk of poor outcomes at the time of diagnosis will make it possible to better choose alternative or experimental therapies. TCF3 (also called E2A) is a transcription factor required in B-lymphopoiesis. Gain-of-function somatic mutations that alter TCF3 or its regulatory partners are associated withBurkitt lymphoma (BL) but not DLBCL. These published findings suggest that biologically-defined, clinically-important subtypes of mature B-cell-derived lymphomas may be distinguished from one another based on differential expression of genes regulated by a master regulator of B-lymphoid biology, namely TCF3. Therefore, in the current study, we hypothesized that biologically- and clinically-important subtypes of DLBCL are distinguishable based upon differential expression of genes that are regulated in B-cells by TCF3. Methods: Fifty-nine subjects who presented with de novo DLBCL at Kingston General Hospital from 2001 to 2010 were identified for inclusion based on the availability of clinical data and sufficient biopsy material for extraction of adequate RNA. We used publically-available microarray-based gene expression and clinical data to identify 99 candidate TCF3 target genes whose differential expression was associated with 3-year overall survival following R-CHOP therapy; these transcripts were represented in aNanoStringCodeSet. Total RNA was purified from cores harvested from representative areas of diagnostic, pre-treatment, formaldehyde-fixed, paraffin-embedded (FFPE) biopsy samples. Twenty-five transcripts whose abundance, individually, was associated with 3-year overall survival in our own cohort were chosen for further study. Results: Unsupervised hierarchical cluster analysis based on relative expression of the 25 TCF3 target genes resolved all except two cases into distinct clusters, denoted A and B, that contained 21 and 36 cases, respectively (heat map at left). In comparing these clusters, Cluster B was associated with rearrangement of MYC (p=0.02) as determined by FISH; no statistically significant associations were evident with age, sex, IPI score, B-symptoms, bulky disease or expression of MYC or BCL2 as determined by immunohistochemistry. Kaplan Meyer (KM) analysis revealed strikingly inferior overall and event-free survival (p=0.0005 for each) for Cluster A cases (KM curve at right). Conclusions: Our findings demonstrate that differential expression of TCF3 target genes ascertained in pre-treatment FFPE biopsy samples of de novo DLBCL defines biologically distinctive subsets of cases with distinctive clinical outcomes following R-CHOP therapy. If validated in an independent cohort, these results may inform both clinical management and the development of new targeted therapies for high-risk cases. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures No relevant conflicts of interest to declare.

Deletion of cIAP1 and cIAP2 in murine B lymphocytes constitutively activates cell survival pathways and inactivates the germinal center response

Blood ◽  
2011 ◽  
Vol 117 (15) ◽  
pp. 4041-4051 ◽  
Author(s):  
Sandra Gardam ◽  
Vivian M. Turner ◽  
Holly Anderton ◽  
Sandhya Limaye ◽  
Antony Basten ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Cell Survival ◽  
Ring Finger ◽  
In Vivo Model ◽  
Cell Physiology ◽  
Ring Finger Proteins ◽  
B Lineage

Abstract B cells require signals delivered through B-cell activating factor of the TNF family receptor (BAFF-R) and CD40 to survive and produce antibody responses in vivo. In vitro data indicate that these signals are controlled by the homologous RING finger proteins cIAP1 and cIAP2, in collaboration with TRAF2 and TRAF3. There is also mounting evidence that all 4 of these signaling molecules can act as tumor suppressors in human B-lineage malignancies. However, it has not been possible to identify the roles of cIAP1 and cIAP2 in controlling B-cell physiology because of the absence of an appropriate in vivo model. Here we describe a unique genetically modified mouse in which the linked cIap1 and cIap2 genes can be independently inactivated. Deletion of cIAP1 plus cIAP2 (but not either protein alone) rendered primary B cells independent of BAFF-R for their survival and led to their uncontrolled accumulation in vivo. B cells deficient in cIAP1 and cIAP2 were also incapable of forming germinal centers, a key step in antibody-mediated immunity. These data define a fundamental role for cIAP1/cIAP2 in regulating B-cell survival and responsiveness, show this requires direct binding to TRAF2, and suggest how mutations of TRAF2, TRAF3, and cIAP1/cIAP2 contribute to B-lineage malignancies, such as multiple myeloma.

scholarly journals Inactivation of CREBBP expands the germinal center B cell compartment, down-regulates MHCII expression and promotes DLBCL growth

2017 ◽  
Vol 114 (36) ◽  
pp. 9701-9706 ◽  
Author(s):  
Hind Hashwah ◽  
Corina A. Schmid ◽  
Sabrina Kasser ◽  
Katrin Bertram ◽  
Anna Stelling ◽  
...  
Keyword(s):  
B Cell ◽  
Germinal Center ◽  
B Cell Lymphoma ◽  
Surface Expression ◽  
Creb Binding Protein ◽  
Cell Compartment ◽  
Genes Encoding ◽  
Additional Cell ◽  
Mhcii Expression ◽  
Serial Transplantation

The genes encoding the histone acetyl-transferases (HATs) CREB binding protein (CREBBP) and EP300 are recurrently mutated in the activated B cell-like and germinal center (GC) B cell-like subtypes of diffuse large B cell lymphoma (DLBCL). Here, we introduced a patient mutation into a human DLBCL cell line using CRISPR and deleted Crebbp and Ep300 in the GC B cell compartment of mice. CREBBP-mutant DLBCL clones exhibited reduced histone H3 acetylation, expressed significantly less MHCII, and grew faster than wild-type clones in s.c. and orthotopic xenograft models. Mice lacking Crebbp in GC B cells exhibited hyperproliferation of their GC compartment upon immunization, had reduced MHCII surface expression on GC cells, and developed accelerated MYC-driven lymphomas. Ep300 inactivation reproduced some, but not all, consequences of Crebbp inactivation. MHCII deficiency phenocopied the effects of CREBBP loss in spontaneous and serial transplantation models of MYC-driven lymphomagenesis, supporting the idea that the mutational inactivation of CREBBP promotes immune evasion. Indeed, the depletion of CD4+ T cells greatly facilitated the engraftment of lymphoma cells in serial transplantation models. In summary, we provide evidence that both HATs are bona fide tumor suppressors that control MHCII expression and promote tumor immune control; mutational inactivation of CREBBP, but not of EP300, has additional cell-intrinsic engraftment and growth-promoting effects.

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