Nodal Lymphoma Research Articles

Primary Cutaneous Lymphomas at A Moroccan Referral Hospital

Primary cutaneous lymphomas (PCL) are the second most frequent form of extra- nodal lymphomas with a broad clinical, histological, phenotypic, genetic, and prognostic spectrum. The aim of this study was to describe the epidemiological, clinical, histopathological, immunohistochemical funding, treatment outcomes and follow up in patients with PCL within Moroccan population.

Follicular Helper T-Cell-derived Nodal Lymphomas: Study of Histomorphologic, Immunophenotypic, Clinical, and RHOA G17V Mutational Profile.

The study was designed to review the demographic, clinical, and pathologic characteristics of follicular helper T cells (TFH)-derived nodal PTCL in India including angioimmunoblastic T-cell lymphoma (AITL), peripheral T-cell lymphoma (PTCL) with follicular helper T cell phenotype (P-TFH), and follicular T-cell lymphoma with additional immunohistochemistry (IHC) and RHOAG17V mutational analysis, as well as their impact on survival. This retrospective study included 88 cases of PTCL that were reclassified using IHC for TFH markers (PD1, ICOS, BCL6, and CD10) and dendritic-meshwork markers (CD21, CD23). Cases of TFH cell origin were evaluated for RHOAG17V mutation using Sanger sequencing and amplification-refractory mutation system-polymerase chain reaction (PCR) (validated using cloning and quantitative PCR) with detailed clinicopathologic correlation. Extensive re-evaluation with added IHC panel resulted in a total of 19 cases being reclassified, and the final subtypes were AITL (37 cases, 42%), PTCL-not otherwise specified (44, 50%), P-TFH (6, 7%), and follicular T-cell lymphoma (1, 1%). The presence of at least 2 TFH markers (>20% immunopositivity) determined the TFH origin. AITL patients tended to be male and showed increased presence of B-symptoms and hepatosplenomegaly. Histomorphology revealed that 92% of AITL cases had pattern 3 involvement. Sanger sequencing with conventional PCR did not yield any mutation, while RHOAG17V was detected by amplification-refractory mutation system-PCR in AITL (51%, P =0.027) and P-TFH (17%), which was validated with cloning followed by sequencing. Cases of RHOAG17V-mutant AITL had a worse Eastern Cooperative Oncology Group performance status initially but fared better in terms of overall outcome ( P =0.029). Although not specific for AITL, RHOAG17V mutation shows an association with diagnosis and requires sensitive methods for detection due to low-tumor burden. The mutant status of AITL could have prognostic implications and translational relevance.

CLINICO-PATHOLOGICAL EXAMINATION OF WALDEYER RING LYMPHOMAS

Introduction: Waldeyer's ring (WR) is the second most common site of extranodal lymphomas. Its clinical course may resemble those of nodal lymphomas. In this study, we evaluated the histopathological and clinical findings, the presence of Epstein-Barr virus (EBV), and overall survival of WR lymphomas diagnosed in our department. Materials and Methods: Retrospectively, 32 patients diagnosed with WR lymphoma between 2017-2022 were included. The demographic, clinical data of the patients and lymphoma types according to the latest classification of the World Health Organization, the presence of EBV (EBV early RNAs (EBER) by chromogenic in-situ hybridization (ISH) method) were examined. Descriptive statistics, categorical data were evaluated as percentage and frequency. Kaplan-Meier analysis was used for survival rate. A P value less than 0.05 was considered statistically significant. Results: The majority of 32 cases in our study were male. The mean age was 57.97±16.15 years. The clinical prediagnosis was carcinoma in 46.9% of the cases. Tonsil and nasopharynx had equal proportions as lymphoma origin. Only 3 (9.4%) cases are classic hodgkin lymphoma (CHL). Others were NHL, of which 16 (50.0%) were diffuse large B-cell lymphoma (DLBCL). EBER was positive only in tonsillar CHL cases. The survival of patients with lymphoma of nasopharyngeal origin was shorter than those of tonsillar origin (p=0.037). Conclusion: WR lymphomas are most often confused with carcinomas clinically. DLBCL is the most common type of lymphoma. EBER has been detected only in CHL. Survival is lower in lymphomas originating from the nasopharynx.
 Keywords: Non-Hodgkin Lymphoma, Nasopharynx, Tonsil, Hodgkin Lymphoma

Tyrosine kinases in nodal peripheral T-cell lymphomas.

Nodal peripheral T-cell lymphomas (PTCL) are uncommon and heterogeneous tumors characterized by a dismal prognosis. Targeted therapy has been proposed. However, reliable targets are mostly represented by a few surface antigens (e.g., CD52 and CD30), chemokine receptors (e.g., CCR4), and epigenetic gene expression regulation. In the last two decades, however, several studies have supported the idea that tyrosine kinase (TK) deregulation might be relevant for both the pathogenesis and treatment of PTCL. Indeed, they can be expressed or activated as a consequence of their involvement in genetic lesions, such as translocations, or by ligand overexpression. The most striking example is ALK in anaplastic large-cell lymphomas (ALCL). ALK activity is necessary to support cell proliferation and survival, and its inhibition leads to cell death. Notably, STAT3 was found to be the main downstream ALK effector. Other TKs are consistently expressed and active in PTCLs, such as PDGFRA, and members of the T-cell receptor signaling family, such as SYK. Notably, as in the case of ALK, STAT proteins have emerged as key downstream factors for most of the involved TK.

IVIM-DKI with parametric reconstruction method for lymph node evaluation and characterization in lymphoma: A preliminary study comparison with FDG-PET/CT

To investigate IVIM-DKI in diagnosing benign and malignant lymph nodes in lymphoma and its subtypes in comparison to FDG-PET/CT. A total of twenty-one (n = 21) patients diagnosed with biopsy-proven Hodgkin lymphoma(HL: n = 13) and non-Hodgkin lymphoma (NHL: n = 8) were prospectively evaluated. All patients underwent MRI(T1-weighted, Short-Tau-Inversion-Recovery (STIR)), and IVIM-DKI was acquired with 9b-values (0–2000s/mm2) at 1.5T and whole-body FDG-PET/CT. The maximum and average standard uptake values (SUVmax and SUVmean) were calculated using PET images. IVIM-DKI parameters (diffusion coefficient(D), pseudo-diffusion coefficient (D*), perfusion fraction (f), and kurtosis (k)) were estimated using IVIM-DKI model with total variation (TV) method (IDTV model). Area-under-curve from receiver-operating-curve (ROC) analysis was used to examine diagnostic value of IVIM-DKI parameters in differentiation between benign and malignant lymph nodes and HL from NHL. Machine learning-based classification of histogram features were performed using linear classifier model. For malignant vs. benign lymph nodes, apparent diffusion coefficient (ADC), f, and k were significantly (p < 0.05) lower in malignant vs. benign lymph nodes. f (AUC:0.88) and k (AUC:0.83) showed high AUC and histogram features combination of f (variance + skewness + kurtosis) showed highest accuracy of 97.2% and AUC of 1. ADC, D, D*, and f were significantly (p < 0.05) lower in NHL than HL. D* showed highest AUC of 0.85 than D (AUC:0.84), ADC (AUC:0.84), and f (AUC:0.74) in NHL vs. HL. SUVmax (spearman-rho = 0.85), ADC (spearman-rho = 0.50), D (spearman-rho = 0.48), and D* (spearman-rho = 0.49) showed significant (p < 0.05) positive correlation with SUVmean. Multi-b-values IVIM-DKI with histogram analysis helps characterize benign and malignant lymph nodes in lymphomas. IVIM-DKI parameters can differentiate malignant lymph nodes in HL and NHL superior to PET parameters.

PET-CT Diagnostic and Evaluation Role in Patients with Nodal Lymphoma

Background: The most prevalent hematological malignancy, malignant lymphoma, makes up around 8% of all adult malignancies. Non-Hodgkin lymphoma (NHL) and Hodgkin disorder (HD) are two types of lymphoid neoplasms. Aim of the Work: Examine how PET/CT may be used to diagnose nodal lymphoma. Analyze the effectiveness of (18 FDG PET/CT) in the first staging of nodal lymphoma. Analyze the function of PET-CT in patient monitoring both during and after the therapeutic course. Examine the function of PET-CT in foretelling and spotting relapses. Patients and Methods: Thirty patients (12 female and 18 males; 11 HD, 19 NHD) with histologically confirmed malignant lymphoma were enrolled in this investigation. They were split into three groups judging by the outcomes of the PET/CT examination. Results: CT for diagnosis was compared to PET/CT results. The current study demonstrated the critical relevance of PET/CT in lymphoma staging, monitoring therapeutic response, and follow-up. The present investigation demonstrated the significance of 18F-FDG. When it comes to the early staging of lymphomas, PET/CT has shown to be a valuable tool in the treatment of lymphomas. PET/CT exhibited substantial effects on early and delayed therapy response evaluation. Conclusion: The outcomes of the present research highlighted the value of 18F- FDG PET/CT as a technique for the first staging of lymphomas. Comparing PET/CT to diagnostic CT has major consequences for early and delayed evaluation of therapy response.

Immunohistochemical analysis of arachidonate 5-lipoxygenase expression in B-cell lymphomas: Implication for B cell differentiation and its analogy with lymphomagenesis.

Arachidonate 5-lipoxygenase (ALOX5) is a cardinal enzyme in the synthesis of leukotrienes, which are powerful immune-regulating lipid mediators. We previously reported that ALOX5 is preferentially expressed in B lymphocytes in the mantle zone of human lymphoid tissue. In the context of physiological relevance, the loss of the Alox5 gene in mice significantly impairs the development of follicular B helper T cells and antibody production. However, ALOX5 expression in B-cell lymphomas has not been investigated in detail. In this study, we examined ALOX5 expression in representative B-cell lymphomas and non-neoplastic lymphoid tissues by immunohistochemistry with a commercially available anti-ALOX5 antibody that can be used on formalin-fixed paraffin-embedded specimens. Interestingly, 22/22 cases of mantle cell lymphoma, 7/7 cases of chronic lymphocytic leukemia/small cell lymphoma, and 20/20 cases of follicular lymphoma expressed ALOX5. A small proportion of extranodal marginal zone lymphoma/mucosa-associated lymphoid tissue lymphoma or nodal marginal zone lymphoma cases were positive for ALOX5 (2/13 or 1/3, respectively). In contrast, no cases with diffuse large B-cell lymphoma, regardless of germinal center B cell (GCB) or non-GCB type, expressed ALOX5 (0/25 cases). These findings suggest that ALOX5 may be a novel marker for identifying the cell of origin of B-cell lymphoma. Further investigation is required to clarify the biological significance of ALOX5 expression in lymphoma cells.

Pathologic and molecular insights in nodal T-follicular helper cell lymphomas.

T-follicular helper (TFH) cells are one of the T-cell subsets with a critical role in the regulation of germinal center (GC) reactions. TFH cells contribute to the positive selection of GC B-cells and promote plasma cell differentiation and antibody production. TFH cells express a unique phenotype characterized by PD-1hi, ICOShi, CD40Lhi, CD95hi, CTLAhi, CCR7lo, and CXCR5hi . Three main subtypes of nodal TFH lymphomas have been described: 1) angioimmunoblastic-type, 2) follicular-type, and 3) not otherwise specified (NOS). The diagnosis of these neoplasms can be challenging, and it is rendered based on a combination of clinical, laboratory, histopathologic, immunophenotypic, and molecular findings. The markers most frequently used to identify a TFH immunophenotype in paraffin-embedded tissue sections include PD-1, CXCL13, CXCR5, ICOS, BCL6, and CD10. These neoplasms feature a characteristic and similar, but not identical, mutational landscape with mutations in epigenetic modifiers (TET2, DNMT3A, IDH2), RHOA, and T-cell receptor signaling genes. Here, we briefly review the biology of TFH cells and present a summary of the current pathologic, molecular, and genetic features of nodal lymphomas. We want to highlight the importance of performing a consistent panel of TFH immunostains and mutational studies in TCLs to identify TFH lymphomas.

Comparison of histological and molecular features of pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma.

Pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma are pediatric B cell lymphomas with similar clinical characteristics but distinct histological features. We investigated the differences between pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma by comparing their histological and molecular characteristics. A total of 5 pediatric-type follicular lymphoma and 11 pediatric nodal marginal zone lymphoma patients were included in the study. In the histological review, 5 of the 16 cases showed overlapping morphological features of pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma; hence, they were reclassified as "mixed type." In molecular analysis, using panel-based massively parallel sequencing, MAP2K1, TNFRSF14, and IRF8 mutations were found in 6, 3, and 2 of the 11 pediatric nodal marginal zone lymphoma patients, respectively, and IRF8 mutation was found in one of the five pediatric-type follicular lymphoma patients. There were no significant differences in genetic alterations established from the histologically reclassified diagnosis as well as the initial diagnosis. Pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma showed morphological overlap in some cases, and no difference between the two was found upon molecular analysis. These findings suggest the possibility that pediatric-type follicular lymphoma and pediatric nodal marginal zone lymphoma are single entity pediatric B-cell lymphoma with broad morphological spectrum.

Combined Reverse-Transcriptase Multiplex Ligation-Dependent Probe Amplification and Next-Generation Sequencing Analyses to Assign Unclassified BCL2−/BCL6− Nonrearranged Small B-Cell Lymphoid Neoplasms as Follicular or Nodal Marginal Zone Lymphoma

Distinguishing between follicular lymphoma (FL) and nodal marginal zone lymphoma (NMZL) can be difficult when morphologic and phenotypic features are unusual and characteristic cytogenetic rearrangements are absent. We evaluated the diagnostic contribution of ancillary techniques—including fluorescence in situ hybridization (FISH)-detected 1p36 deletion; reverse-transcriptase, multiplex, ligation-dependent probe amplification (RT-MLPA); and next-generation sequencing (NGS)—for tumors that remain unclassified according to standard criteria. After review, 50 CD5-negative small B-cell lymphoid neoplasms without BCL2 and BCL6 FISH rearrangements were diagnosed as FLs (n = 27), NMZLs (n = 5), or unclassified (n = 18) based on the 2016 World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues. FISH helped identify the 1p36 deletion in 3 FLs and 1 unclassified tumor. Most classified FLs had an RT-MLPA germinal center B-cell (GCB) signature (93%) or were noncontributive (7%). Classified NMZLs had an RT-MLPA activated B-cell signature (20%), had an unassigned signature (40%), or were noncontributive (40%). Among unclassified tumors, the RT-MLPA GCB signature was associated with mutations most commonly found in FLs (CREBBP, EZH2, STAT6, and/or TNFRSF14) (90%). An RT-MLPA-detected GCB signature and/or NGS-detected gene mutations were considered as FL identifiers for 13 tumors. An activated B-cell signature or NOTCH2 mutation supported NMZL diagnosis in 3 tumors. Combining the RT-MLPA and NGS findings successfully discriminated 89% of unclassified tumors in favor of one or the other diagnosis. NGS-detected mutations may be of therapeutic interest. Herein, we detected 3 EZH2 and 8 CREBBP mutations that might be eligible for targeted therapies.

Clinicopathological Characteristics and Prognosis of Diffuse Large B-Cell Lymphoma in Relation to CA-125 and CA 19-9 Expression.

Background: Some epithelial tumors express the carbohydrate antigen 125 (Cancer antigen-125, CA-125) and CA 19-9, especially ovarian and pancreatic tumors. Patients with non-Hodgkin lymphoma (NHL) were reported to have a close association between serum CA-125 levels and adverse prognostic factors with worse survival. We aimed to investigate CA-125 and 19-9 expression in nodal diffuse large B-cell lymphoma, not otherwise specified (DLBCL NOS) tissues using immunohistochemistry (IHC) and their relations to clinicopathological manifestations and patients' survival. Methods: 65 cases of DLBCL NOS were examined. A modified mechanical pencil tip was used to construct Manual Tissue Micro-array (TMA) blocks. Immunohistochemical staining for CA-125 and CA 19-9 was performed and scored semi-quantitatively. All relations were analyzed using established statistical methodologies. Results: Aberrant expression of CA 19-9 was detected in 12% of cases without any expression of CA-125. Moreover, 75% of the CA 19-9 positive cases were statistically significantly associated with anemia and performance status 1. Also, 75% of the CA 19-9 positive cases were females. Conclusions: CA 19-9 was aberrantly expressed in 12% of nodal DLBCL NOS cases and significantly related to anaemia and performance status but not to survival. In cases of DLBCL NOS, CA 19-9 expression cannot be considered an independent prognostic factor. CA-125 was not expressed in nodal DLBCL NOS tissues, necessitating re-evaluation studies. Therefore, it is advised to conduct more research to clarify the potential correlation between serum and tissue CA 19-9 levels and other clinic-pathological characteristics of nodal and extranodal DLBCL NOS patients.

Radiological Assessment of Retroperitoneal Neoplastic Lesions in a Tertiary Care Hospital, Rajkot, Gujarat, India: A Cross-sectional Study

Introduction: The retroperitoneum is an important potential space in the human abdomen. Retroperitoneal (RP) tumours are extremely vast in position, size, rate of growth and in pathological types. They silently grow before giving rise to symptoms or become palpable. Radiology plays a unique role in sustaining the optimum information for the diagnosis of lesions of retroperitoneal space. Computed Tomography (CT) is generally more superior in the diagnosis of retroperitoneal neoplastic lesions in comparison to X-Ray or Ultrasonography (USG) but later are cost-effective and readily available. Aim: To describe the clinical spectrum of retroperitonial neoplastic lesions and also to evaluate the radiographic features of different retroperitoneal neoplastic lesions with various radiological modalities. Materials and Methods: This cross-sectional study was conducted on 50 patients from November 2019 to April 2021, in Department of Radiology of Pandit Deendayal Upadhyay Government Medical College and Civil Hospital, Rajkot, Gujarat. Each patient had undergone CT, USG and X-ray as indicated. Descriptive data was analysed and presented in terms of frequencies and percentages. Results: In this study among 50 patients, males (60%) were found to be affected more than the females (40%) with a male:female ratio of 1.5:1. The most common malignant RP neoplastic lesion was metastatic nodal masses and lymphoma. 41 (82%) of cases were malignant while 9 (18%) were benign neoplasms. The most common benign RP neoplastic lesion was teratoma. The most common presentation in patients with retroperitoneal lesions was pain and lump in the abdomen with abdominal distension. Most of the malignant lesions were of average size more than 10 cm. Conclusion: Most of the retroperitoneal neoplastic lesions were malignant and seen in males. CT, USG and X-ray are helpful in the characterisation and diagnosis of retroperitoneal neoplastic masses.

Intravascular Large B-Cell Lymphoma (IVLBCL) Presenting with CNS Involvement in Patient with Chronic Lymphocytic Leukemia

Background: Intravascular lymphoma (IVL) is a rare, often fatal disease characterized by intraluminal proliferation of lymphoid cells within blood vessels. Intravascular Large B-Cell Lymphoma (IVLBCL) is frequently found in different organs, but the skin and central nervous system (CNS) are the most affected ones. Routine radiological examination and additional diagnostic tools, such as high-resolution, three-dimensional (3D) FLAIR sequence, T2-weighted (T2W) and T1-weighted (T1W) contrast-enhanced magnetic resonance imaging (MRI), MRI diffusion and perfusion imaging studies are useful to confirm the diagnosis in cases where it is not easy to perform histopathological examinations [1]. 1.2. Case Report: We present a clinical case of a 63-year-old female patient who presented with progressive cognitive deficit as mild motor aphasia, acalculia, agraphia, dyslexia and mild ideomotor apraxia without any signs of other organ involvement. It was known that the patient had 5-year anamnesis of chronic lymphocytic leukemia, stage II and splanchnic venous thrombosis 2 months ago. Blood biochemical analysis without any significant change, except leukocytosis in full blood count (leukocytes 15,2 x 10*9/L, where neutrophils were in normal range, but lymphocytes were increased). Head magnetic resonance imaging (MRI) showed pathological signal areas on the left parietal lobe with hypointense signal on contrast-enhanced T1-weighted, hyperintense signal on contrast-enhanced T2-weighted and FLAIR-weighted MRI images with restricted diffusion and slightly lower ADC value. Magnetic resonance perfusion imaging studies did not show an increase in cerebral blood flow or in cerebral blood volume. In this case, the patient’s diagnosis was made based on typical radiological features. 1.3. Conclusions: Intravascular large B-cell lymphoma is a rare subtype of extra nodal large B-cell lymphoma with an aggressive clinical course. The diagnosis is challenging, as it presents without any obvious tumour mass or lymphadenopathy. [2] As the clinical presentation is variable without specific characteristics, mostly biopsy or even autopsy confirms the definite diagnosis. But in some cases, the diagnosis can be made by typical radiological findings in the restricted availability of brain biopsy.

Crystal-Storing Histiocytosis in a Patient with Nodal Marginal Zone Lymphoma

We present a case of a 72-year-old male diagnosed with concurrent crystal-storing histiocytosis and nodal marginal zone lymphoma with plasmacytic differentiation following a finding of new axillary lymphadenopathy during active surveillance for an early stage lung cancer.

Severe tumor lysis syndrome after rituximab-bendamustine in nodal marginal zone lymphoma

Marjinal zon lenfomalar, Hodgkin dışı lenfomalar arasında en sık görülen üçüncü alt tiptir. Nodal marjinal zon lenfoma, marjinal zon lenfomanın alt tiplerinden biridir. Genellikle altıncı dekattan sonra ortaya çıkar. Tüm lenfomalar içinde yaklaşık %1 görülme oranına sahip, B hücreli bir lenfomadır. Nodal marjinal zon lenfoma tedavisinde, tümör lizis sendromu (TLS) nadir görülmektedir. Aynı zamanda herhangi bir nedenle rituksimab-bendamustin kemoterapisinde de TLS beklenti değildir. Olgumuz, nodal marjinal zon lenfoma tanılı hastada rituximab-bendamustin kemoterapi tedavisi sonrası bulantı ve kusma şikayetleri ile kliniğimize başvurmuş olup TLS saptanmıştır. Sonuç olarak hastalar düşük dereceli lenfoma bile olsa dev kitlesi ve LDH yüksekliğinde özellikle akut böbrek hasarı tablosu ile başvurduğunda TLS olasılığı göz önünde bulundurulmalıdır.

Distinct and overlapping features of nodal peripheral T-cell lymphomas exhibiting a follicular helper T-cell phenotype: a multicenter study emphasizing the clinicopathological significance of follicular helper T-cell marker expression

Nodal peripheral T-cell lymphoma (PTCL) is a heterogeneous category including angioimmunoblastic T-cell lymphoma (AITL), PTCL of follicular helper T-cell (Tfh) phenotype (PTCL-Tfh), and PTCL, not otherwise specified (PTCL-NOS). We explored Tfh marker profiles in nodal PTCL. Nodal PTCLs (n=129) were reclassified into AITL (58%; 75/129), PTCL-Tfh (26%; 34/129), and PTCL-NOS (16%; 20/129). Histologically, clear cell clusters, high endothelial venules, follicular dendritic cell proliferation, EBV+cells, and Hodgkin-Reed-Sternberg (HRS)-like cells were more common in AITL than PTCL-Tfh (HRS-like cells, P=.005; otherwise, P<.001) and PTCL-NOS (HRS-like cells, P=.028; otherwise, P<.001). PTCL-NOS had a higher Ki-67 index than AITL (P=.001) and PTCL-Tfh (P=.002). Clinically, AITL had frequent B symptoms (versus PTCL-Tfh, P=.010), while PTCL-NOS exhibited low stage (versus AITL+PTCL-Tfh, P=.036). Positive Tfh markers were greater in AITL (3.5±1.1) than PTCL-Tfh (2.9±0.9; P=.006) and PTCL-NOS (0.5±0.5; P<.001). Tfh markers showed close correlations among them and AITL-defining histology. By clustering analysis, AITL and PTCL-NOS were relatively exclusively clustered, while PTCL-Tfh overlapped with them. Survival was not different among the PTCL entities. By Cox regression, sex and ECOG performance status (PS) independently predicted shorter progression-free survival in the whole cohort (male, P=.001, HR=2.5; PS≥2, P=.010, HR=1.9) and in 'Tfh-lymphomas' (ie, AITL+PTCL-Tfh) (male, P=.001, HR=2.6; PS≥2, P=.016, HR=2.1), while only PS predicted shorter overall survival (OS) in the whole cohort (P=.012, HR=2.7) and in 'Tfh-lymphomas' (P=.001; HR=3.2). ICOS predicted favorable OS in 'Tfh-lymphomas' (log-rank; P=.016). Despite the overlapping features, nodal PTCL entities could be characterized by Tfh markers revealing clinicopathologic implications.

Development and validation of nomograms by radiomic features on ultrasound imaging for predicting overall survival in patients with primary nodal diffuse large B-cell lymphoma.

To develop and validate a nomogram to predict the overall survival (OS) of patients with primary nodal diffuse large B-cell lymphoma(N-DLBCL) based on radiomic features and clinical features. A retrospective analysis was performed on 145 patients confirmed with N-DLBCL and they were randomly assigned to training set(n=78), internal validation set(n=33), external validation set(n=34). First, a clinical model (model 1) was established according to clinical features and ultrasound (US) results. Then, based on the radiomics features extracted from conventional ultrasound images, a radiomic signature was constructed (model 2), and the radiomics score (Rad-Score) was calculated. Finally, a comprehensive model was established (model 3) combined with Rad-score and clinical features. Receiver operating characteristic (ROC) curves were employed to evaluate the performance of model 1, model 2 and model 3. Based on model 3, we plotted a nomogram. Calibration curves were used to test the effectiveness of the nomogram, and decision curve analysis (DCA) was used to asset the nomogram in clinical use. According to multivariate analysis, 3 clinical features and Rad-score were finally selected to construct the model 3, which showed better predictive value for OS in patients with N-DLBCL than mode 1 and model 2 in training (AUC,0. 891 vs. 0.779 vs.0.756), internal validation (AUC, 0.868 vs. 0.713, vs.0.756) and external validation (AUC, 914 vs. 0.866, vs.0.789) sets. Decision curve analysis demonstrated that the nomogram based on model 3 was more clinically useful than the other two models. The developed nomogram is a useful tool for precisely analyzing the prognosis of N-DLBCL patients, which could help clinicians in making personalized survival predictions and assessing individualized clinical options.

S78 Diffuse Large B Cell Lymphoma of the Anal Transition Zone After Ileal Pouch Anal Anastomosis: A Report of 4 Cases

Background: Development of lymphoma at the anal transition zone (ATZ) following restorative proctocolectomy with ileal pouch anal anastomosis (IPAA) is extremely rare, with less than 10 cases described in the literature. Methods: We retrospectively analyzed our prospectively collected database on pelvic pouches. Patients who developed lymphoma at the ATZ were included in this study, and oncological and functional outcomes were described. Results: Out of 5,070 patients undergoing IPAA between 1983 and 2021, 4 developed lymphoma of the ATZ (3 males, 1 female). The histology was diffuse large B cell lymphoma (DLBCL) in all cases. All patients had a diagnosis of ulcerative colitis, with concomitant primary sclerosing cholangitis (PSC) in 2 cases. The indication to IPAA was colon cancer in 2 cases, dysplasia in one case, and fulminant colitis in one case. Three patients had a J pouch, while one had an S pouch. The median interval between IPAA and lymphoma development was 7 years (range 3-12). Two patients had a previous history of hematologic malignancies (acute myeloid leukemia and nodal marginal zone lymphoma, respectively), while one was immunosuppressed due to liver transplantion for PSC. In 3 cases, the ATZ was the first site involved, while one case first presented as a thyroid lymphoma, achieved remission with R-CHOP, and later developed a recurrence at the ATZ. Three patients had stage IAE DLBCL at presentation, while one patient had stage IV disease. In all cases, the presenting symptoms were akin to pouchitis, with no patient exhibiting B symptoms. Two patients had concomitant acute kidney injury due to lymphoma-related obstructive uropathy with bilateral hydronephrosis requiring bilateral nephrostomies. All patients received R-CHOP as first line treatment, with all patients with stage IAE disease achieving remission. The patient with stage IV DLBCL was one of the 2 patients with obstructive uropathy: he was diverted with loop ileostomy and died of lymphoma-related sepsis 3 months after lymphoma development. Similarly, the other patient with obstructive uropathy developed a recurrence shortly after remission and died of lymphoma-related sepsis 10 months after lymphoma development. Regarding the other 2 patients, the post-transplant patient developed recurrence 3 years after remission: immunosuppression was stopped, and she went into remission one month later. She had no further recurrences, and died 4 years later due to progressive multifocal leukoencepalopathy with her pouch still intact. The last patient, who had no predisposing factors for lymphoma development, had no recurrence; he is still alive with no evidence of disease and his pouch intact 7.5 year after lymphoma development. Conclusion(s): Diffuse large B cell lymphoma of the ATZ is a rare occurrence that can be effectively treated with chemotherapy without resulting in pouch failure. Although uncommon, lymphoma should be considered in the differential in patients with symptoms of pouchitis and predisposing factors or a history of hematologic malignancies.

Follicle Center Lymphoma (FCL) of the Lower Female Genital Tract (LFGT): A Novel Variant of Primary Cutaneous Follicle Center Lymphoma (PCFCL).

Primary cutaneous follicle center lymphoma has been distinguished from nodal follicular lymphoma (FL) based on genomic and clinical features. The nature of other extranodal FLs is not well defined. We report 15 cases of follicle center lymphoma involving the lower female genital tract. Cases were evaluated using an immunohistochemical panel for B-cell lymphoma, B-cell clonality, fluorescence in situ hybridization for BCL2 gene rearrangement, and next-generation sequencing. All patients had localized disease with no evidence of bone marrow involvement. Most cases (12/15, 80%) had a follicular pattern, at least focally. Large centrocytes were a prominent feature leading to concern for diffuse large B-cell lymphoma by referring pathologists. Neoplastic cells were positive for CD20 and BCL-6, while BCL-2 was positive in 2/15 (13%) cases. Fluorescence in situ hybridization for BCL2 gene rearrangement was negative in 10/11 (91%) cases. Next-generation sequencing performed in 10 cases revealed TNFRSF14 as the most frequently mutated gene in 6/10 (60%) cases. No case had CREBBP or KMT2D mutations as seen in nodal FL. None of the patients had progressive disease with durable complete remission achieved in 10/12 (83%) cases. The median follow-up period was 7.8 years (range: 0.2 to 20.5y) with a 5-year overall survival of 100%. We conclude that follicle center lymphoma of the lower female genital tract is a novel variant of primary cutaneous follicle center lymphoma. Despite a frequent component of large cells, it is characterized by localized disease and low risk for dissemination. Awareness and recognition are important to distinguish these lesions from aggressive B-cell lymphomas.

Clinical Development of Senza5TM CART5: a Novel Dual Population CD5 CRISPR-Cas9 Knocked out Anti-CD5 Chimeric Antigen Receptor T Cell Product for Relapsed and Refractory CD5+ Nodal T-Cell Lymphomas

Introduction: Chimeric antigen receptor T-cell (CART) immunotherapy is FDA-approved for B-cell lymphoblastic leukemia, B-cell non-Hodgkin lymphomas, and multiple myeloma. However, CART immunotherapy for T-cell non-Hodgkin lymphomas (T-NHL) is lagging, with only limited clinical results. T-NHL have a poor prognosis and there is an unmet need for effective and long-lasting treatments. We previously presented preliminary preclinical results of anti-CD5 CAR T cells (CART5) that had CD5 knocked out (KO) (Blood (2020) 136 (Supplement 1): 51-52.). CD5 KO not only prevented CART fratricide but also dramatically enhanced CART function. Critical challenges to developing CART cells for T-NHL are CART fratricide and normal T-cell toxicity. In this study, we addressed these challenges by improving manufacturing and including a dual-population technology to mitigate toxicity. We present the development of Senza5TM CART5 an autologous good-manufacturing practice (GMP)-grade product (Fig.1) that will be tested in a phase I clinical trial for relapsed and refractory CD5+ nodal T-cell lymphomas. Methods and Results: We first identified a single guide RNA (sgRNA) that deletes CD5 in T cells with >90% KO efficiency by flow cytometry and ICE analysis. We then assessed the specificity of the KO by performing iGUIDE sequencing. We confirmed high on-target CD5 cleavage with minimal off-targets (CALCP, C20orf85, INPP4B, XPO7, and SLC10A7). These genes have little to no expression in naïve CD4+ and CD8+ T cells by RNA-seq (DICE). Targeted functional assays are ongoing. For the CAR construct, we selected a lead CAR5 among six 4-1BB-constructs based on its superior anti-tumor function in NOD-SCID IL2Rgnull (NSG) mice engrafted with a Jurkat T-lymphoblastic leukemia cell line. Furthermore, to confirm the specificity of the CAR5 for CD5, we screened CAR5 T cells for interactions with HEK293 cells expressing 5,828 human plasma membrane proteins or cell-surface tethered secreted proteins and 398 human heterodimers (Retrogenix assay). The lead CAR5 did not bind to any targets besides CD5. For an optimized GMP-grade product, we modified our manufacturing protocol, shortening it from 10 to 5 days. Moreover, KO efficiency and safety were optimized to reduce off-target activity without loss of efficiency. We also optimized the activation strategy to decrease cell loss. Functional studies were performed to demonstrate the comparability of these methods. With these improvements, we were able to achieve >90% CD5 KO with >30% CAR transduction, thereby allowing the generation of two main populations: one of CD5KO CART5 cells and the other of CD5KO normal untransduced T cells. CD5KO CART5 are expected to target CD5+ T-NHL cells but could also lead to normal T-cell toxicity. This toxicity will be mitigated by the CD5KO normal T cells that will have a survival advantage after infusion (Fig.1) and have been demonstrated to maintain reactivity against opportunistic pathogens such as CMV and EBV. To evaluate Senza5TMCART5 safety, we performed cytokine-independent CART proliferation and long- and short-term xenograft models. Senza5TM CART5 cells did not grow aberrantly without growth/survival-associated cytokines (IL2/7/15). We then used NSG mice to exclude uncontrolled Senza5TM CART5 growth and observed enhanced expansion and proliferation of Senza5TM CART5 with a xenogeneic GVHD rate similar to that of control mice with similar wild-type CART5 expansion. Since the CAR5 does not cross-react with murine CD5, we tested non-specific toxicity in tissues using an NSG xenograft model of Jurkat. On Day 10 post-CAR T injections, animals were euthanized, and a complete necropsy was performed on all animals. Senza5TM CART5 demonstrated tumor clearance with no detection of treatment-related lesions. Mice treated with untransduced T cells or CART19 demonstrated significant overall and organ-specific weight loss compared to Senza5TM CART5. Blood analyses determined no differences in their clinical chemistry profile or complete blood counts. Conclusion: In this study, we developed a novel dual-population, full-scale GMP-ready CD5 KO anti-CD5 CART product for T-cell lymphomas characterized by enhanced anti-tumor effect and a strategy to potentially mitigate toxicity. Autologous Senza5TM CART5 will be tested for CD5+ nodal T-cell lymphomas in a phase I clinical trial. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal