Figure 2 - uploaded by Frank Vari
Content may be subject to copyright.
![Mutational landscape of PCNSL, according to EBV tissue and HIV serological status. (A) Each column in this plot represents an individual case (with mutation[s] in the displayed genes) of the final sequencing cohort (n 5 79), across the 4 tissue subtypes: EBV 2 HIV 2 PCNSL, EBV 2 HIV 1 PCNSL, EBV 1 HIV 1 PCNSL, and EBV 1 HIV 2 PCNSL. Mutated genes constitute individual rows and are sorted according to their mutational frequencies of mutated cases as provided on the far right. Mutation types are color coded as indicated in the key; red* brain lymphoma TK cell-line, red** patients without PTLD/iatrogenic immunosuppression. (B) Stacked histograms show the percentage (percentages rounded to whole numbers) of cases with mutations in MYD88, CD79B, PIM1 by EBV-tissue and HIV serological status across the 3 main subtypes. Because EBV 2 HIV 1 PCNSL represented only 1 sequenced case, aggregate data are not shown. (C) Number of mutated genes observed using the targeted sequencing panel in EBV 2 HIV 2 PCNSL, EBV 1 HIV 1 PCNSL, and EBV 1 HIV 2 PCNSL, with P values for paired subtypes: *P # .05; **P # .01; ***P # .001; ****P # .0001.](profile/Frank-Vari/publication/347004192/figure/fig1/AS:1025250930225154@1621450278922/Mutational-landscape-of-PCNSL-according-to-EBV-tissue-and-HIV-serological-status-A.png)
Mutational landscape of PCNSL, according to EBV tissue and HIV serological status. (A) Each column in this plot represents an individual case (with mutation[s] in the displayed genes) of the final sequencing cohort (n 5 79), across the 4 tissue subtypes: EBV 2 HIV 2 PCNSL, EBV 2 HIV 1 PCNSL, EBV 1 HIV 1 PCNSL, and EBV 1 HIV 2 PCNSL. Mutated genes constitute individual rows and are sorted according to their mutational frequencies of mutated cases as provided on the far right. Mutation types are color coded as indicated in the key; red* brain lymphoma TK cell-line, red** patients without PTLD/iatrogenic immunosuppression. (B) Stacked histograms show the percentage (percentages rounded to whole numbers) of cases with mutations in MYD88, CD79B, PIM1 by EBV-tissue and HIV serological status across the 3 main subtypes. Because EBV 2 HIV 1 PCNSL represented only 1 sequenced case, aggregate data are not shown. (C) Number of mutated genes observed using the targeted sequencing panel in EBV 2 HIV 2 PCNSL, EBV 1 HIV 1 PCNSL, and EBV 1 HIV 2 PCNSL, with P values for paired subtypes: *P # .05; **P # .01; ***P # .001; ****P # .0001.
Source publication
Primary central nervous system lymphoma (PCNSL) is confined to the brain, eyes, and cerebrospinal fluid without evidence of systemic spread. Rarely, PCNSL occurs in the context of immunosuppression, e.g. post-transplant lymphoproliferative disorders (PTLD) or HIV (AIDS-related PCNSL). These cases are poorly characterized, have dismal outcome and ar...
Contexts in source publication
Context 1
... of the mutational landscapes across PCNSL subtypes Figure 2A shows results of individual samples grouped by PCNSL subtypes. First, EBV tissue-negative PCNSL: EBV 2 HIV 2 PCNSL and EBV 2 HIV 1 PCNSL cases were analyzed. ...
Context 2
... we examined the frequency with which MYD88, CD79B, and PIM1 mutations cooccurred ( Figure 2B). These 3 mutations are contained in the recently proposed "C5" and "MCD" DLBCL molecular classifications. ...
Context 3
... total number of mutated genes detected in the targeted panel were next compared ( Figure 2C). There was a higher number of mutations observed in EBV 2 HIV 2 PCNSL compared with EBV 1 HIV 1 PCNSL and EBV 1 HIV 2 PCNSL cases (P 5 2.0 3 10 29 , and P 5 3.7 3 10 215 , respectively). ...
Context 4
... of the mutational landscapes across PCNSL subtypes Figure 2A shows results of individual samples grouped by PCNSL subtypes. First, EBV tissue-negative PCNSL: EBV 2 HIV 2 PCNSL and EBV 2 HIV 1 PCNSL cases were analyzed. ...
Context 5
... we examined the frequency with which MYD88, CD79B, and PIM1 mutations cooccurred ( Figure 2B). These 3 mutations are contained in the recently proposed "C5" and "MCD" DLBCL molecular classifications. ...
Citations
... Immunosuppression is a well-known risk factor for PCNSL although found in a minority of patient [2][3][4][5][6][7][8][9]. These lymphomas represent a distinct histopathological and molecular entity driven by EBV oncogenesis in most cases [10][11][12][13][14]. ...
... Moreover, 28% of the lesions were hemorrhagic at baseline, as described in prior studies [25,[29][30][31]33]. These differences in tumor nature could be explained by a different genetic and immunological background found in EBV positive PCNSL [9,14], though data was missing in this cohort to confirm this hypothesis. ...
Purpose
Immunosuppression is a well-established risk factor for primary central nervous system lymphomas (PCNSLs), which present in this context distinct radiological characteristics. Our aim was to describe the radiological evolution of treated PCNSL in immunocompromised patients and suggest adapted MRI response criteria.
Methods
We conducted a multicenter retrospective study of patients from the French LOC, K-Virogref and CANCERVIH network databases and enrolled adult immunocompromised patients with newly diagnosed PCNSL.
Results
We evaluated the baseline, intermediate, end-of-treatment and follow-up MRI data of 31 patients (9 living with HIV, 16 with solid organ transplantation and 6 with an autoimmune disease under chronic immunosuppressive therapy). At baseline, 23/30 (77%) patients had necrotic lesions with ring enhancement and 28% of the lesions were hemorrhagic. At the end of the first-line treatment, 12/28 (43%) patients could not be classified according to the IPCG criteria. Thirteen of 28 (46%) patients still harbored contrast enhancement, and 11/28 (39%) patients had persistent large necrotic lesions with a median diameter of 15 mm. These aspects were not associated with a pejorative outcome and progressively diminished during follow-up. Six patients relapsed; however, we failed to identify any neuroimaging risk factors on the end-of-treatment MRI.
Conclusion
In immunocompromised patients, PCNSLs often harbor alarming features on end-of-treatment MRI, with persistent contrast-enhanced lesions frequently observed. However, these aspects seemed to be related to the necrotic and hemorrhagic nature of the lesions and were not predictive of a pejorative outcome. Specific response criteria for this population are thereby proposed.
... These findings are consistent with previous studies. 4,25,29 Advancements in understanding the pathogenesis of NHLs associated with oncogenic viruses could provide valuable treatment targets. ...
Objective
Immune tolerance and evasion play a critical role in virus‐driven malignancies. However, the phenotype and clinical significance of programmed cell death 1 (PD‐1) and its ligands, PD‐L1 and PD‐L2, in aggressive acquired immunodeficiency syndrome (AIDS)‐related non‐Hodgkin lymphoma (AR‐NHL) remain poorly understood, particularly in the Epstein–Barr virus (EBV)‐positive subset.
Methods
We used in situ hybridization with EBV‐encoded RNA (EBER) to assess the EBV status. We performed immunohistochemistry and flow cytometry analysis to evaluate components of the PD‐1/PD‐L1/L2 pathway in a multi‐institutional cohort of 58 patients with AR‐NHL and compared EBV‐positive and EBV‐negative cases.
Results
The prevalence of EBV ⁺ in AR‐NHL was 56.9% and was associated with a marked increase in the expression of PD‐1/PD‐L1/PD‐L2 in malignant cells. Patients with AR‐NHLs who tested positive for both EBER and PD‐1 exhibited lower survival rates compared to those negative for these markers (47.4% vs. 93.8%, p = 0.004). Similarly, patients positive for both EBER and PD‐L1 also demonstrated poorer survival (56.5% vs. 93.8%, p = 0.043). Importantly, PD‐1 tissue‐expression demonstrated independent prognostic significance for overall survival in multivariate analysis and was correlated to elevated levels of LDH ( r = 0.313, p = 0.031), increased PD‐1 ⁺ Tregs ( p = 0.006), and robust expression of EBER ( r = 0.541, p < 0.001) and PD‐L1 ( r = 0.354, p = 0.014) expression.
Conclusions
These data emphasize the importance of PD‐1‐mediated immune evasion in the complex landscape of immune oncology in AR‐NHL co‐infected with EBV, and contribute to the diagnostic classification and possible definition of immunotherapeutic strategies for this unique subgroup.
... The incidence of PCNSL in individuals infected with human immunode ciency virus (HIV) ranges from 2-6%, which is 1,000 times greater than that in HIV-negative individuals [10]. More than 96% of PCNSLs are highly invasive B-cell lymphomas, with DLBCL being the most common subtype, while T-cell and low-grade lymphomas account for 1-4% of PCNSLs [11,12]. In 2004, three antibodies (CD10, Bcl-6, and Mum-1) were utilized by ...
Background AIDS-related primary central nervous system lymphoma (AR-PCNSL) differs from immunocompetent-primary central nervous system lymphoma (IC-PCNSL) in certain features. The main objective of this study was to investigate the differences in clinicopathological features between AR-PCNSL and IC-PCNSL.
Methods Thirty-seven AR-PCNSL patients and thirty IC-PCNSL patients were included. hematoxylin & eosin staining; immunohistochemical detection using CD20, Bcl-2, Bcl-6, p53, C-MYC, Ki67, and METTL3 antibodies; and Epstein–Barr encoding region (EBER) in situ hybridization were performed.
Results All of the observed patients were classified as the DLBCL histological type. AR-PCNSL were younger (37.7 vs. 60.5 years) and had a higher likelihood of being male (86.5% vs. 63.3%) than non-AIDS patients were. Elevated LDH and low sugar content in cerebrospinal fluid (CSF) were more common among AR-PCNSL. The expression levels of METTL3, Bcl-2 and p53 expressions were significantly higher in AR-PCNSL patients than in PCNSL patients without AIDS. In contrast, AR-PCNSL patients exhibited lower levels of Bcl-6 expression. AR-PCNSL patients were more likely to be positive for EBER, accounting for 81.1% of these patients. Furthermore, we also found that the expression of METTL3 was lower in GCB-like DLBCL (n=7) than in ABC-like DLBCL (n=32) in AR-PCNSL (p=0.041); however, in IC-PCNSL patients, the expression of METTL3 was not significantly different between GCB-like DLBCL and ABC-like DLBCL (p=0.710).
Conclusions Our study of Chinese AR-PCNSL and IC-PCNSL patients revealed new findings, indicating that METTL3, Bcl-2 and p53 were increased in AR-PCNSL patients compared to IC-PCNSL patients and that METTL3 was higher in ABC-like DLBCL patients than in GCB-like DLBCL in AR-PCNSL patients, suggesting a notable distinction in the pathological characteristics between PCNSL patients with or without AIDS.
... 58 EBV-associated PCNSL appears to represent a unique biological entity characterised by retention of antigen presentation (in contrast to EBV-negative PCNSL) and a tolerogenic TME comprising overexpression of the inhibitory cell surface molecules PDL-1, PDL-2, LAG3 and TIM-3. 73 The prognosis for patients with PTLD involving the CNS has been shown to be inferior to that of other PCNSL patients 74 but appears to be better for those with isolated CNS involvement and may have improved in the modern era. 58 Treatment of these patients tends to involve combinations of radiotherapy, chemotherapy and rituximab, with recent evidence showing potential for improved outcomes with both modifications of existing regimens to involve higher doses of methotrexate, 74 and with novel approaches more directed to the underlying biology, such as BTK inhibition. ...
Epstein–Barr virus (EBV)‐associated lymphomas cover a range of histological B‐ and T‐cell non‐Hodgkin and Hodgkin lymphoma subtypes. The role of EBV on B‐cell malignant pathogenesis and its impact on the tumour microenvironment are intriguing but incompletely understood. Both the International Consensus Classification (ICC) and 5th Edition of the World Health Organization (WHO‐HAEM5) proposals give prominence to the distinct clinical, prognostic, genetic and tumour microenvironmental features of EBV in lymphoproliferative disorders. There have been major advances in our biological understanding, in how to harness features of EBV and its host immune response for targeted therapy, and in using EBV as a method to monitor disease response. In this article, we showcase the latest developments and how they may be integrated to stimulate new and innovative approaches for further lines of investigation and therapy.
... 10,11 Although, a more recent comprehensive study based on gene expression analysis in primary central nervous system lymphomas suggests that EBV association with ABC DLBCL may be lower. 12 Interestingly, EBNA2 expression is also associated with drug resistance in DLBCL. 13 Worldwide, over 95% of the human population is EBV seropositive. ...
Hematological malignancies like Burkitt lymphoma (BL), Hodgkin's lymphoma (HL) and diffuse large B cell lymphoma (DLBCL) cause significant morbidity in humans. A substantial number of these lymphomas, particularly, HL and DLBCLs have poorer prognosis due to their association with Epstein-Barr virus (EBV). Our earlier studies have shown that EBV encoded nuclear antigen EBNA2 upregulates PD-L1 in DLBCL and BLs by downregulating miR-34a. Here, we investigated whether EBNA2 affects the inducible co-stimulator ligand, ICOSL, a molecule required for efficient recognition of tumor cells by T cells through the engagement of ICOS on the latter. In virus-infected and EBNA2-transfected B-lymphoma cells, ICOSL expression was reduced. Our investigation of the molecular mechanisms revealed that this was due to an increase in miR-24 by EBNA2. By using ICOSL 3'UTR-Luc reporter system, we validated that ICOSL is an authentic miR-24 target. Transfection of anti-miR-24 molecules in EBNA2 expressing lymphoma cells reconstituted ICOSL expression and increased tumor immunogenicity in mixed lymphocyte reactions. Since miR-24 is known to target c-MYC, an oncoprotein positively regulated by EBNA2, we analyzed its expression in anti-miR-24 transfected lymphoma cells. Indeed, the reduction of miR-24 in EBNA2 expressing DLBCL further elevated c-MYC and increased apoptosis. Consistent with the in vitro data, EBNA2 positive DLBCL biopsies expressed low ICOSL and high miR-24. We suggest that EBV evades host immune responses through EBNA2 by inducing miR-24 to reduce ICOSL expression and for simultaneous rheostatic maintenance of pro-proliferative c-MYC levels. Overall, these data identify miR-24 as potential therapeutically relevant target in EBV associated lymphomas.
... Gandhi et al studied the genetics of PCNSL in immune deficient patients, most associated with EBV infection.70 In contrast to the genetic profile of immune competent patients, almost all cases lacked mutation of MYD88 and CD79B.Moreover, most cases retained surface HLA expression. ...
Diffuse large B-cell lymphoma (DLBCL) is an aggressive B-cell lymphoma curable even in advanced stages. DLBCL involving the central nervous system (CNS) is more difficult to cure and fewer treatment options exist. Primary CNS lymphoma (PCNSL) refers to aggressive lymphomas confined to the CNS, and are almost always DLBCL. Standard approaches for PCNSL use high-dose methotrexate based combinations as induction therapy and younger patients often receive dose-intensive consolidation. However, dose-intensive therapies are not suitable for all patients, and older patients have fewer effective treatment options. Patients with relapsed or chemotherapy-refractory disease have a grave prognosis. Secondary CNS lymphoma (SCNSL) describes aggressive lymphomas involving the CNS at initial presentation or relapses within the CNS after treatment for systemic DLBCL. Isolated CNS relapse is often managed as PCNSL, but patients with synchronous involvement of DLBCL in both the periphery and the CNS pose a unique clinical challenge. Insights into the molecular circuitry of DLBCL have identified distinct genetic subtypes including cases with a predilection for CNS invasion. PCNSL and subsets of SCNSL are characterised by chronically activated B-cell receptor and NFKB signalling along with genetic evidence of immune evasion which may be exploited therapeutically. Improved mechanistic understanding of targetable pathways underpinning CNS lymphomas has led to numerous clinical trials testing targeted agent combinations and immunotherapy approaches with promising early results. Biologically rational strategies may further improve the cure rate of CNS lymphomas either by overcoming intrinsic or acquired treatment resistance and/or by being broadly applicable to patients of all ages.
... In our study, 7.2% (9/124) of patients were EBV positive (detected by EBV-encoded small RNAs or serum EBV DNA), but due to a lack of testing in most patients (97 unknown), the data is less reliable. EBV-associated PCNSL in immunosuppressed patients is immunobiologically distinct from EBV-negative and HIV-negative PCNSL [19]. Restoration of EBV-specific T-cell immunity has been shown to induce a clinical response in EBV-positive lymphomas [19,20]. ...
... EBV-associated PCNSL in immunosuppressed patients is immunobiologically distinct from EBV-negative and HIV-negative PCNSL [19]. Restoration of EBV-specific T-cell immunity has been shown to induce a clinical response in EBV-positive lymphomas [19,20]. Hence, the role of EBV in the development of PCNSL warrants further investigation in clinical practice. ...
To explore prognostic factors and outcomes of primary central nervous system lymphoma (PCNSL) of diffuse large B-cell lymphoma (DLBCL) in Taiwan, 124 PCNSL-DLBCL patients (from 1995 to 2021) were retrospectively analyzed. Mainly, two treatment modalities including sandwich chemoradiotherapy and modified MATRix regimen were employed in these patients. Overall survival (OS) was determined by log-rank test and time-dependent Cox analysis. Median OS of all patients was 27.1 months. 47 (37.9%) patients who underwent sandwich chemoradiotherapy had a complete remission (CR) rate of 87.2%, median OS of 53.9 months, and progression free survival (PFS) of 42.9 months. 11 (8.9%) patients who underwent modified MATRix regimen had CR rate of 72.7%, median OS of 18.9, and PFS of 11.2 months. There are no significant OS differences between treatment groups or addition of Rituximab. Patients treated with the modified MATRix regimen experienced a higher early mortality rate followed by a survival plateau. IELSG low-risk group had significantly improved OS and PFS than IELSG intermediate- or high-risk group. In multivariant analysis, age > 60 years old and bilateral cerebral lesions are associated with significantly inferior OS. Sandwich chemoradiotherapy demonstrated better early survival and reduced treatment-related toxicity for PCNSL patients compared to the modified MATRix regimen. However, the long-term follow-up revealed a higher rate of treatment failure events in the sandwich chemoradiotherapy group. IELSG and MSKCC scores served as reliable risk assessment models. Incorporating bilateral cerebral lesions as a risk factor further improved risk evaluation.
... Necrosis and high mitotic activity are present. The lymphoma cells are intermingled with reactive bystander cells consisting of small mature T and B lymphocytes, macrophages, activated microglial cells, and reactive astrocytes [228]. In addition to mature B-cell antigens, CD20, CD22, CD79a, CD19, and PAX5, the lymphoma cells express IRF4/MUM1, BCL2, BCL6, and IGM, have a high (>80-90%) Ki67 proliferation index, and are negative for CD38 and CD138 [12]. ...
The diagnosis and treatment of lymphoid neoplasms have undergone a continuously progressive positive change in the last three decades, with accelerated progress in the previous decade due to the advent of genomics in cancer diagnosis. Significantly, there has been an increasing emphasis on integrating molecular genetics with clinical, morphologic, immunophenotypic, and cytogenetic evaluation for diagnosis. As we think of moving forward with further advances in the genomics era, it is first helpful to understand our current state of knowledge and how we achieved it in the challenging and complex field of lymphoid neoplasms, which comprise very heterogeneous neoplastic diseases in children and adults, including clinically acute lymphoblastic leukemias (ALLs) arising from precursor lymphoid cells and clinically indolent and aggressive lymphomas arising from mature lymphoid cells. This work aims to provide an overview of the historical evolution and the current state of knowledge to anyone interested in the field of lymphoid neoplasms, including students, physicians, and researchers. Therefore, I discuss this complex topic in three review manuscripts, designated Parts 1–3. In Part 1, I explain the basis of the diagnostic classification of lymphoid neoplasms and its evolution up to the current fifth edition of the World Health Organization classification of hematolymphoid neoplasms, and the crucial importance of diagnostic tumor classifications in achieving and advancing patient care and precision medicine. In the second and third manuscripts, I discuss current diagnostic considerations for B-ALL and T-ALL (Part 2) and common indolent and aggressive mature leukemias/lymphomas (Part 3), including significant updates in the WHO 2022 classification, newly described entities, and concepts, including genetic predisposition to ALLs and lymphomas, and throughout emphasizing the essential integration of molecular genetics with clinical, morphologic (pathologic), immunophenotypic, and cytogenetic evaluation, as is required for precise diagnosis of the type of lymphoma/leukemia in any patient.
... 6,55 Glioblastoma, metastases, and opportunistic infections are the primary differential. 57,[62][63][64][65] PTLD PTLD is a rare EBV-driven polyclonal B-cell proliferation occurring in patients posttransplantation. CNS involvement is seen in 5%-20% of PTLDs after solid organ transplantation and portends a poor prognosis. ...
Epstein-Barr virus is a ubiquitous herpesvirus that may cause both infective (encephalitis, meningitis, and so forth) and postinfection inflammatory (such as Guillain-Barré syndrome, acute disseminated encephalomyelitis) manifestations in the CNS. Diagnosis of Epstein-Barr virus-related CNS pathologies is often complicated due to a nonspecific clinical presentation and overlap with other infectious and noninfectious causes, both clinically and on imaging. The Epstein-Barr virus is also implicated in several lymphoproliferative disorders in both immunocompromised and immunocompetent hosts. MR imaging is preferred for evaluating the extent of involvement and monitoring therapy response, given its high sensitivity and specificity, though imaging findings may be nonspecific. Herein, we review the imaging spectrum of Epstein-Barr virus-associated CNS disorders.
... Interestingly, upregulation of proteins that are recognized by the B-cells is observed in reaction of inflammation and tumoral infiltration in the CNS compartment such as endoglin and galectine-3 expressed by the CNS endothelial cells [12]. By contrast, in immunodeficient patient, the molecular pathogenesis is specifically different, as immunodeficiency-associated lymphomas are EBV-related [20]. ...
Purpose of review:
Primary central nervous system lymphoma (PCNSL) is a rare central nervous system (CNS) malignancy, which represents a heterogenous group of tumors. Among PCNSL, diffuse large B-cell lymphoma of the CNS (CNS-DLBCL) represents the most common tumor type. Multiomics studies have recently revealed the complex genomic landscape of these rare diseases. These findings lead to a potential new molecular and epigenetic classification.
Recent findings:
Our review is focused on CNS-DLBCL in immunocompetent patients. CNS-DLBCL are derived from self-reactive/polyreactive precursor cells. An early molecular event such as MYD88 mutation leads to escape elimination of precursor cells, which, by a dysregulated GC reaction, acquire auto-/polyreactivity of the B-cell tumoral cells for antigens physiologically expressed in the CNS. Most of CNS-DLBCL tumor cells harbor a non-GCB, ABC-like immunophenotype associated with a late GC (exit) B-cells genotype by gene expression profiling. Various mechanisms of genetic alterations are involved in the pathogenesis of PCNSL, including point mutations [nonsomatic hypermutation (SHM), aberrant SHM (aSHM)], SHM/aSHM, chromosome copy gains or losses, and DNA hypermethylation. Constitutive NFκB activation plays a key role in lymphoma cell proliferation and survival by dysregulation of toll-like receptor (mutations of CARD11 and MYD88), BCR (CD79B), JAK-STAT, and NFκB signaling pathways.
Summary:
Multiomics approaches have succeeded to substantially improve the understanding of the pathogenesis, as well as the molecular and epigenetic events in PCNSL. Challenges remain due to the obvious heterogeneity of CNS-DLBCL, and improvement is needed for their classification.
