Diagnosis Of Cutaneous T-cell Lymphoma Research Articles

Micro-RNAs in the Diagnosis of Cutaneous T-Cell Lymphomas

Rationale. Early diagnosis of mycosis fungoides (MF), as the most common form of T-cell lymphoma, presents significant challenges. The diagnosis of MF is based on the following criteria: comprehensive assessment of the clinical picture of the disease, histological and immunohistochemical examination of the skin, and determination of rearrangement of the T-cell receptor gene, but even this does not always aid in diagnosis. The aim of the study is to investigate the expression of micro-RNAs (miR-223, -423, -663, -16, -326, -711) in the blood plasma and leukocytes of patients with a presumptive diagnosis of MF to improve the disease diagnosis. Methods. This study included 50 patients aged 24 to 79 years, of whom 30 patients had a preliminary diagnosis of MF and 20 patients with small plaque parapsoriasis, who formed the comparison group. All patients underwent histological, immunohistochemical examination of skin biopsies, and determination of micro-RNA (miR-223, -423, -663, -16, -326, -711) expression in blood plasma and leukocytes by real-time PCR. Results. Analyzing the results of clinical-anamnestic, histological, and immunohistochemical research methods, the diagnosis of MF was established in 22 of 30 (73.3%) patients, of which 9 of 14 (64.3%) were in the early stages of the disease. small plaque parapsoriasis. Conclusion. During our study, it was found that the studied micro-RNAs (miR-326, -663, -711, -223, -423, -16) in the blood plasma and leukocytes of patients with MF have statistically significant levels of expression compared to the low level of expression of these micro-RNAs in patients with small plaque parapsoriasis. The expression of micro-RNAs we studied in the skin contributes to the improvement of MF diagnosis with an accuracy of up to 90%.

Application of Blood Lymphocyte Immunophenotype and TCR Gene Rearrangement in the Diagnosis of CTCL.

The immunophenotype of peripheral blood lymphocytes and T-cell receptor (TCR) gene rearrangement of cutaneous T cell lymphoma (CTCL) patients were retrospectively analyzed to explore their value in the diagnosis of CTCL. A total of fifty patients' results were enrolled from 2013 to 2021, including 29 malignant skin disorders and 21 benign skin disorders. The immunophenotype of peripheral blood lymphocytes were analyzed by flow cytometry and TCR gene rearrangement was detected by capillary electrophoresis. Lymphocyte subsets, CD4/CD8 ratio, the percentage of CD3+CD4+CD7- cells and CD45RA/CD45RO ratio was calculated between malignant and benign skin disorders. Peripheral blood lymphocyte immunophenotype and TCR gene rearrangement was compared with skin biopsy to evaluate their sensitivity and specificity. Lymphocyte subsets between malignant and benign groups have no significant difference in percentage of T cell (p > 0.05). The CD4/CD8 ratio is higher in patients with malignant lymphoma than the healthy range. The percentage of CD3+CD4+CD7- cells in malignant groups is higher than that in benign groups and CD45RA/ CD45RO ratio has significant difference between malignant and benign groups (p < 0.05). The sensitivity and specificity of TCR rearrangement for CTCL were 51.7% and 42.9%. The sensitivity and specificity of peripheral blood lymphocyte immunophenotype for CTCL were 44.8% and 33.3%. Combining the two methods, the sensitivity and specificity reached 69.0% and 38.1%, respectively. CD4/CD8 ratio of lymphocyte subsets, the proportion of CD4+CD7-T cells and CD45RA/CD45RO ratio can effectively distinguish benign and malignant dermatosis. TCR rearrangement method combined with lymphocyte immunophenotype can improve the sensitivity and specificity of CTCL diagnosis.

Cutaneous T-Cell Lymphoma and Dupilumab Use: A Retrospective Matched Cohort Study of Clinical Characteristics and Treatment Outcomes

Introduction: Dupilumab is a human monoclonal antibody that inhibits signaling of IL-4 and IL-13 and is currently approved by the Food and Drug Administration for the treatment of atopic dermatitis (AD). While causality has not been established, sporadic reports have emerged associating dupilumab use with the onset, progression, or subsequent diagnosis of cutaneous T-cell lymphomas, in particular mycoses fungoides (MF). The natural history of MF diagnosed after dupilumab exposure remains unclear. We performed a retrospective matched cohort study to compare the clinical characteristics and treatment outcomes of patients with MF based on prior dupilumab exposure. Methods: Through the Memorial Sloan Kettering (MSK) Research &amp; Technology data delivery platform (DataLine), we conducted a search for any patient seen by a provider in the MSK Cutaneous Lymphoma Clinic whose electronic record contained the word ‘dupilumab’ or ‘Dupixent.‘ Once the initial query was performed, we individually reviewed each patient to confirm: (1) A histologically-confirmed diagnosis of MF by an MSK dermatopathologist, and (2) Use of dupilumab prior to the histological diagnosis of MF. After identifying a cohort of patients with confirmed MF after dupilumab use (cases), we identified a matched cohort of patients with confirmed MF and no prior dupilumab use (controls). Specifically, we matched patients for stage at diagnosis, date of birth and age at diagnosis (+/- 5 years), and date of presentation (+/- 5 years). Clinical data and treatment outcomes were collected by clinical review. Baseline characteristics were summarized using descriptive statistics and compared between the two groups using the Wilcoxon rank sum test, Pearson's Chi-squared test, and Fisher's exact test. Overall survival (OS), progression-free survival (PFS), and recurrence-free survival (RFS) were estimated as the elapsed time from initial diagnosis to death, disease recurrence, and progression, respectively. Kaplan-Meier estimates were stratified by exposure to dupilumab, and differences between the estimates were compared using the log-rank test. Results: We identified 25 patients who presented with a new, histologically-confirmed diagnosis of MF following dupilumab treatment for a diagnosis of AD. In 16 of these patients, AD was diagnosed based on a skin biopsy, whereas in 9 patients the diagnosis was based on clinical features only. Among 16 patients with known duration of dupilumab use, the median duration of use was 5.0 months (min/max: 1.5-24 months). In total, 14 patients were diagnosed with early-stage disease (IA n=2, IB n=12) and 11 were diagnosed with advanced stage disease (IIB n=4, IIIA n=2, IVA1 n=5). The estimated time from dupilumab treatment to lymphoma diagnosis was 10.2 months, and the median age at lymphoma diagnosis was 53 years (min/max: 28-84). Compared to the cohort of 25 matched patients with MF and no prior dupilumab use ( Table 1), no difference was observed in race, sex, initial therapy (skin-directed vs. systemic), or total number of therapies. The median follow-up time for patients exposed to dupilumab was 13.2 months (95% CI: 6.8-21.9), and 20.7 months (95% CI: 13.4-27.8) for those not exposed. No differences were observed in OS, PFS, or RFS between the exposed (dupilumab) and non-exposed (non-dupilumab) cohorts ( Figure 1). Two-year OS, PFS, and RFS in the non-dupilumab cohort was 85.0%, 47.3%, and 70.7%, compared to 100.0%, 41.5%, and 79.6% in the dupilumab cohort. One patient in the dupilumab-exposed cohort died of a non-lymphoma malignancy, whereas 4 patients in the non-exposed cohort died (3 due to lymphoma, 1 due to unknown causes). Conclusions: In summary, we identified a relatively large cohort of patients with MF diagnosed after dupilumab treatment. Patients presented with early and advanced-stage disease. Outcome were comparable to a matched cohort of non-exposed patients with MF. The causal relationship between dupilumab and MF remains unproven and deserves dedicated biological study.

Coexistence of psoriasis and cutaneous T-cell lymphoma.

It has been reported that individuals with psoriasis are at an increased risk of developing cutaneous T-cell lymphoma (CTCL). However, the increased risk of lymphoma in these patients has been questioned because CTCL in its early stages may be incorrectly labelled as psoriasis, thus introducing potential for misclassification bias. We retrospectively reviewed patients with a confirmed diagnosis of CTCL seen in a tertiary cutaneous lymphoma clinic (n = 115) over a 5-year period and found that 6 (5.2%) patients had clinical evidence of coexisting psoriasis. This demonstrates that there is a small cohort of individuals who develop both psoriasis and CTCL.

110 Non-invasive molecular analysis for CTCL detection and differentiation from other skin conditions with adhesive smart sticker skin sample collection

Cutaneous T cell lymphoma (CTCL) is a complex and heterogenous disease with a range of pathogenic, diagnostic, prognostic and therapeutic courses. Broadly, CTCL is characterized by cutaneous infiltrates of aberrant monocolonal T-lymphocytes. Clinically and histopathologically, CTCL often mimics psoriasis, eczema and other benign dermatoses. Diagnosis of CTCL is often delayed by 4-6 years from the time lesions initially appear, and typically requires multiple skin biopsies and blood tests. In this study, we employed non-invasive adhesive patches to collect samples from lesional skin of CTCL (n=12) and psoriasis (n=10) patients as well as non-lesional skin from healthy (n=12) volunteers.

154 Increasing incidence of cutaneous t-cell lymphoma in the United States: A SEER population data analysis

Recent incidence patterns of cutaneous T-cell lymphoma (CTCL) in the United States are not well described. Our goal was to evaluate incidence trends of CTCL by subtype and examine associations to sex, age, race/ethnicity, and socioeconomic status (SES). Using data from the Surveillance, Epidemiology, and End Results (SEER) Registry, we calculated age-adjusted incidence rates and annual percentage change (APC) in incidence of CTCL. We identified 14,942 patients with a new diagnosis of CTCL between 2000 and 2018.

LB936 Cutaneous T-cell lymphoma (CTCL) following B-cell lymphoma: A case series

Cutaneous T-cell lymphoma (CTCL) is associated with an increased risk of developing secondary malignancies, particularly lymphomas. However, the diagnosis of lymphoma prior to CTCL is very rare. Herein we present the cases of two patients diagnosed with lymphoma prior to a diagnosis of CTCL. The first patient was a female in her late 50s with a history of diffuse large B-cell lymphoma (DLBL) of her breast status post chemo plus radiation who developed poikilodermatous scaly patches on her arms and legs one year after her DLBL diagnosis. Her condition was initially managed as an eczema refractory to treatment until a biopsy performed at an outside clinic several years later revealed likely CTCL followed by confirmative biopsies demonstrating poikiloderma atrophicans vasculare. She was treated with acitretin 10 mg five times weekly and narrowband UVB twice weekly for 12 treatments, but self-discontinued both. She recently initiated monthly extracorporeal photopheresis. The second patient was a female in her 70s with history of melanoma who presented to outpatient dermatology for evaluation of erythematous macules on her groin pending radiation treatment for DLBL of her breast. The rash was refractory to topical therapies and a biopsy later demonstrated mycosis fungoides. The patient initially preferred conservative therapy with natural sunlight and topical hydrocortisone, but ultimately elected for monthly photopheresis as her disease progressed. The response of both patients to photopheresis is yet to be determined. These cases illustrate the importance of full workup for erythematous, scaly rashes especially in the setting of other malignancy and in rashes refractory to topical treatments for other conditions. Both cases had at least one biopsy unrevealing of the true underlying CTCL diagnosis, showing that multiple biopsies may be necessary to capture this diagnosis.

Characterization of primary cutaneous T-cell lymphoma following solid organ transplantation.

Immunosuppression following solid organ transplantation is a known risk factor for the development of posttransplant lymphoproliferative disorders (PTLD). Primary cutaneous T-cell lymphoma (CTCL) occurring in the posttransplant setting is rare, which has made comprehensive understanding of this disease challenging. This study aims to further characterize the spectrum of clinicopathologic features of CTCL in solid organ transplant recipients (SOTR). A retrospective chart review was performed for SOTR who were diagnosed with CTCL at a multi-site academic medical center from January 1, 1998, to December 31, 2013. Eight patients fulfilled the inclusion criteria and were included in this study. Data collected included patient demographics, transplanted organ, the time between transplant and CTCL diagnosis, clinical presentation and rash morphology, a histological subtype of CTCL, immunosuppression regimens, and patient status. Twelve diagnostic skin biopsies for five patients were examined and reviewed by a board-certified dermatopathologist. Six (75%) out of the eight patients were men, two (25%) were women, and the median age was 53 years. The median time from the date of transplant to the diagnosis of CTCL was 8.2 years. Transplanted organs included the liver (4), kidney (3), and heart (1). Clinical presentation varied from papulonodules, comedone-like lesions, intense pruritis, and scaly erythematous eruptions. The most common histologic presentation was folliculotropic mycosis fungoides (FMF) (7/12). Epstein-Barr virus-in situ hybridization (EBV-ISH) was negative in all specimens. We emphasize the rarity of CTCL among SOTR. Although rare in the general population, the FMF subtype appears to be disproportionately seen in SOTR compared with other CTCL.

Трудности дифференциальной диагностики грибовидного микоза и псориаза: клинико-морфологические сопоставления

Mycosis fungoides is an epidermotropic primary cutaneous T-cell lymphoma, represented com-posed of small and medium-sized lymphocytes with cerebriform nuclei. Differential diagnosis of cutaneous T-cell lymphoma and chronic autoimmune dermatoses, including psoriasis, may present significant chal-lenges in some cases. We analyzed three clinical cases of mycosis fungoides, in which psoriasis was initially diagnosed. Histological and immunohistochemical studies with a wide panel of antibodies (CD4 CD8, CD2, CD5, CD7, CD3) were performed, and monoclonal T-lymphocyte populations were determined with PCR. We propose an algorithm for diagnosing mycosis fungoides. Keywords: mycosis fungoides, psoriasis, morphology, immunohistochemistrycal study, diagnosis

Modern possibilities of therapy for primary cutaneous T-cell lymphomas: the first results of the use of brentuximab vedotin in the Russian Federation

Background. Primary cutaneous T-cell lymphomas are rare heterogeneous group of lymphoproliferative diseases characterized by primarily involving skin and subcutaneous adipose tissue. Half of these cases are mycosis fungoides (MF), for about 25% are cutaneous CD30+ lymphoproliferative diseases (CD30+ LPD): primary cutaneous anaplastic large cell lymphoma (pcALCL) and lymphomatoid papulosis (LyP). During the initiating treatment of patients with MF and Szary syndrome (SS), carried out on the territory of the Russian Federation, for about 30% of patients are resistant to various therapeutic effects, especially in the later stages. The problem of the treatment of CD30+ LPD is extracutaneous dissemination in case of pcALCL, steadily relapsing course of LyP without symptom-free intervals. These characteristics of the therapy of cutaneous lymphomas demand for the need to search for new treatment options. Brentuximab vedotin, according to the results of the international randomized ALCANZA trial, has shown high efficiency in the treatment of cutaneous T-cell lymphoproliferative diseases. Aim. To evaluate the efficacy of brentuximab vedotin application in patients with cutaneous T-cell lymphomas in adverse risk group received at least one line of systemic therapy. Materials and methods. The study included 21 patients: 16 men and 5 women. The diagnosis of MF was verified in 8 patients, SS in 5 patients, cutaneous CD30+ LPD in 6 patients (5 patients pcALCL, 1 patient LyP) and a primary cutaneous peripheral T-cell lymphoma, unspecified in 2 patients. The diagnosis of cutaneous T-cell lymphoma was verified on the basis of the anamnesis of the disease, on the character of cutaneous lesions, on histological, immunohistochemical and in some cases on molecular genetic testing of the biopted sample of the skin (the assessment of T-cell receptor gene rearrangement). Results. The late stages of the disease were diagnosed in 12 of 13 patients with MF/SS. Extracutaneous lesions were diagnosed in 57% of cases. The median of prior lines therapy was 3 (18 variants of treatment). The overall response to the treatment was achieved in 91% of cases (in 19 of 21 patients): the complete remission was obtained in 53% of cases, very good partial remission in 31% of cases and partial remission in 16% of cases. The progression of the disease was determined in 2 patients (after the first and fourth cycles). Some patients with partial remission as a result of therapy using brentuximab vedotin had the additional therapy (radiation therapy, interferon , the cycles of systemic therapy) and these acts gave an option of achieving deeper antitumor response. The early relapse was diagnosed in 2 of 19 patients who had responded to the treatment. The treatment tolerability was acceptable, and the toxicity did not exceed the already known one described in earlier studies. Thus, the stable overall antitumor response had been persisting in 89% of patients (the median of the observation was 10 months). Conclusion. The use of targeted therapy with brentuximab vedotin gave an option of achieving high treatment results in group of patients with advanced stages of the disease and inefficiency of several lines of therapy.

Response to Extracorporeal Photopheresis in Patients with Cutaneous T-Cell Lymphoma: A Retrospective Medical Chart Review

INTRODUCTIONExtracorporeal photopheresis (ECP) is an immunomodulatory therapy that has been used in the treatment of cutaneous T-cell lymphoma (CTCL) for over 30 years. Clinical trials of newer agents for the treatment of CTCL have shown response rates of approximately 30% in heterogeneous patient populations. The objective of this study was to assess the effectiveness of ECP in the treatment of CTCL patients in real-world clinical practice.METHODSThis is an interim analysis of a retrospective medical chart review study conducted at clinical sites in the United States, in which treating physicians were responsible for patient selection and data collection via structured case report forms. Patients with a confirmed diagnosis of CTCL who initiated ECP between January 1, 2017 and February 28, 2019 were included in the study. In addition, patients must have initiated ECP at age ≥18 years with no ECP treatment received within the year prior to data collection, have received at least 3 months of ECP treatment, and have response data available in the patient chart. Data collected included patient demographics, clinical characteristics, and treatments received prior to and concomitantly with ECP. Clinical outcomes were collected every 3 months during treatment for up to 18 months, and included the body surface area (BSA) affected, appearance of new skin lesions, and the physician-rated Clinical Global Impression-Improvement (CGI-I) scores. Response to ECP was defined as >50% reduction in BSA affected at any point during the follow-up period, consisting of the time from ECP initiation through the time of data collection. Data analysis was descriptive in nature.RESULTSFour clinical sites participated in the study and enrolled a total of 26 patients for the interim analysis. Patients were predominantly female (53.8%) and White (88.5%). Mean age at CTCL diagnosis was 68.4 years, and the majority patients were diagnosed with Sézary syndrome (57.5%) or mycosis fungoides (30.8%). Nearly half of patients (46.1%) had stage IV disease at diagnosis (IVA, 26.9%; IVB, 19.2%) and half of patients (50.0%) had lymph node involvement at diagnosis. Median BSA involvement with plaques/patches at diagnosis was 80% (interquartile range [IQR], 20% to 90%). Six patients (23.1%) achieved >50% reduction in BSA affected, and among those, median time to response was 6.0 months (IQR, 2.9 to 15.0 months). New skin lesions appeared in 5 patients (19.2%). Among those with available data, the percentage of patients rated as minimally improved, much improved, or very much improved on the CGI-I was 57.7% at 3 months (N=26), 50.0% at 6 months (N=18), and 60.0% at 9 months (N=15) after ECP initiation.CONCLUSIONSThis retrospective observational study describes patient characteristics and clinical outcomes among CTCL-diagnosed patients initiating therapy with ECP. Despite the population treated with ECP in real-world practice being older and having more advanced-stage disease compared to recent clinical trials, response rate was comparable. DisclosuresGirardi: Transimmune: Patents & Royalties: Inventor/IP; Helsinn: Membership on an entity's Board of Directors or advisory committees, Research Funding; AbbVie: Research Funding; Soligenix: Research Funding; Mallinckrodt Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; Stradefy Biosciences Inc: Patents & Royalties: Inventor/IP; Actelion: Membership on an entity's Board of Directors or advisory committees, Research Funding. Huang: Mallinckrodt Pharmaceuticals: Current Employment. Corman: Mallinckrodt Pharmaceuticals: Consultancy. Edmundson: Mallinckrodt Pharmaceuticals: Consultancy. Kale: Mallinckrodt Pharmaceuticals: Consultancy. Rusibamayila: Mallinckrodt Pharmaceuticals: Consultancy. Foss: Kura: Honoraria; Daiichi Sankyo: Honoraria; Seattle Genetics: Honoraria, Speakers Bureau; Acrotech: Honoraria, Speakers Bureau; Mallinckrodt: Honoraria; Kyowa: Honoraria.

A Rare and Fatal Case of Cutaneous T-Cell Lymphoma (CTCL)

Introduction: Cutaneous T-Cell Lymphoma (CTCL) is a form of Non-Hodgkin Lymphoma involving T-cell neoplasms mainly concentrated in the skin. Primary Cutaneous Peripheral T-Cell Lymphoma is Not Otherwise Specified (PTL-NOS) is the rarest case of CTCL disease.Case Presentation: A female, 35 years old, complained of a lump on her face. In the last 3 months, the lump felt soft, contained a liquid that, when it ruptures, secretes Blood, and the crusty scar will blacken and easily bleed with pain. Physical examination of pale conjunctiva on the face, neck, chest, abdomen, back, and legs obtained nodules with a chewy consistency with erythema appearance in varying size. Some nodules that appear were covered in blackish crustae. The right lung has a decrease in the sound of breath, accompanied by swelling in both limbs. The conclusion of bilateral femur skin biopsy impresses cutaneous T lymphoma cells. Within three months, cancer developed into lesions spread almost throughout the body, and due to the rapid and progressive nature of cancer, its diagnosis developed into PTL-NOS.Conclusions: Patients with PTL-NOS may come with solitary nodules such as red tumors in any area of the body. However, most often, patients come with symptoms of multifocal or diffuse scattered nodules. In enforcing the diagnosis, it takes a high level of suspicion, and multiple rebiopsies are necessary to enforce the diagnosis of CTCL.

Bio-O1-01 - Diagnostic performance of high throughput sequencing of the T-cell receptor beta gene for the diagnosis of cutaneous T-cell lymphoma

The early diagnosis of MF and SS may be challenging and is based on a combination of clinical, biological, histological, and molecular criteria. Until now, the search for T-cell clones in skin was mostly performed by standard PCR amplification of the T-cell receptor (TCR) genes and capillary electrophoresis (CE) but false positives and negatives with this technique hamper MF diagnosis. High throughput sequencing (HTS) of the TCR genes allows the identification and quantification of each T-cell clone through the recognition of CDR3 sequences in tissue samples. It can be used for the sensitive monitoring of the minimal residual disease in CTCL. However, the interpretation of HTS clonality data remains subjective, as internationally validated thresholds for the definition of a dominant T-cell clone by HTS have not been published. In this study we sought to evaluate the diagnostic performance and define optimal thresholds for T-cell clonality of HTS of TCR beta gene (HTS-TRB) in CTCL diagnosis. We sought to evaluate the diagnostic performance and define optimal thresholds for T-cell clonality of HTS of TCR beta gene (HTS-TRB) by analyzing the TCF, ratio of productive frequencies (RPF, defined as the ratio between the 1st and 2nd clone frequencies) and clonality score in patients with CTCL compared to inflammatory skin diseases (ID). We retrospectively analyzed HTS-TRB data of lesional skin biopsy samples from 144 patients between 2013 and 2020. CTCL diagnosis was based on international clinical, histopathological and immunohistochemical criteria. Data were analyzed using SPSS software. A total of 101 samples from patients with certain CTCL, of which 47 MF stage I–IIA, 1 stage IIB, 4 stage III, 35 SS (3), 8 peripheral T-cell lymphomas not otherwise specified (PTCL NOS) or other aggressive epidermotropic CD8-positive T-cell lymphoma and 6 CD30+ lymphoproliferative disorders (3 lymphomatoid papulosis and 3 anaplastic large-cell lymphomas), were collected. In addition, 43 samples from patients with ID were identified. The performance of the HTS-TRB technique for CTCL diagnosis was analyzed using receiver-operating characteristics (ROC) curves on 101 CTCL and 43 ID cases. The TCF showed the highest diagnostic performance with an area under curve of 0.957 versus 0.929 for RPF. With 5% and 25% TCF thresholds, the specificity for CTCL diagnosis was 95% and 100%, and sensitivity 89% and 50%, respectively. Such a high specificity allows to early identify CTCL in difficult cases. Further multicentre prospective studies are needed to validate international criteria for T-cell clonality analysis by HTS of TCR genes in the diagnosis of CTCL. The early diagnosis of MF and SS may be challenging and is based on a combination of clinical, biological, histological, and molecular criteria. Until now, the search for T-cell clones in skin was mostly performed by standard PCR amplification of the T-cell receptor (TCR) genes and capillary electrophoresis (CE) but false positives and negatives with this technique hamper MF diagnosis. High throughput sequencing (HTS) of the TCR genes allows the identification and quantification of each T-cell clone through the recognition of CDR3 sequences in tissue samples. It can be used for the sensitive monitoring of the minimal residual disease in CTCL. However, the interpretation of HTS clonality data remains subjective, as internationally validated thresholds for the definition of a dominant T-cell clone by HTS have not been published. In this study we sought to evaluate the diagnostic performance and define optimal thresholds for T-cell clonality of HTS of TCR beta gene (HTS-TRB) in CTCL diagnosis. We sought to evaluate the diagnostic performance and define optimal thresholds for T-cell clonality of HTS of TCR beta gene (HTS-TRB) by analyzing the TCF, ratio of productive frequencies (RPF, defined as the ratio between the 1st and 2nd clone frequencies) and clonality score in patients with CTCL compared to inflammatory skin diseases (ID). We retrospectively analyzed HTS-TRB data of lesional skin biopsy samples from 144 patients between 2013 and 2020. CTCL diagnosis was based on international clinical, histopathological and immunohistochemical criteria. Data were analyzed using SPSS software. A total of 101 samples from patients with certain CTCL, of which 47 MF stage I–IIA, 1 stage IIB, 4 stage III, 35 SS (3), 8 peripheral T-cell lymphomas not otherwise specified (PTCL NOS) or other aggressive epidermotropic CD8-positive T-cell lymphoma and 6 CD30+ lymphoproliferative disorders (3 lymphomatoid papulosis and 3 anaplastic large-cell lymphomas), were collected. In addition, 43 samples from patients with ID were identified. The performance of the HTS-TRB technique for CTCL diagnosis was analyzed using receiver-operating characteristics (ROC) curves on 101 CTCL and 43 ID cases. The TCF showed the highest diagnostic performance with an area under curve of 0.957 versus 0.929 for RPF. With 5% and 25% TCF thresholds, the specificity for CTCL diagnosis was 95% and 100%, and sensitivity 89% and 50%, respectively. Such a high specificity allows to early identify CTCL in difficult cases. Further multicentre prospective studies are needed to validate international criteria for T-cell clonality analysis by HTS of TCR genes in the diagnosis of CTCL.

Assessment of Clonotypic Rearrangements and Minimal Residual Disease in Lymphoid Malignancies.

Measurable (minimal) residual disease (MRD) is an independent prognostic factor for survival outcomes in patients with lymphoid and plasma cell malignancies and has been incorporated into consensus criteria regarding treatment response, strategy, and clinical trial endpoints. clonoSEQ (a next-generation sequencing [NGS]-MRD assay) uses multiplex polymerase chain reaction and NGS to identify clonotypic rearrangements at the immunoglobulin (Ig) H, IgK, IgL, T-cell receptor (TCR)-β, and TCR-γ loci, as well as translocated B-cell lymphoma 1/IgH and 2/IgH sequences for MRD assessment. Additionally, it can be used to confirm diagnoses of cutaneous T-cell lymphoma (CTCL). To review the technical aspects of our experience using the clonoSEQ Assay in routine clinical practice. In this single-center experience, 390 patients with lymphoid and plasma cell malignancies were assessed with the NGS-MRD Assay at a central laboratory. Median time from arrival of the shipment to initiation of the assay (defined as captured in Adaptive's secure tracking system) was 2.1 hours. Overall, 317 patients had 1 or more samples submitted for sequence identification. Of these, 290 (91.5%) had trackable sequences identified. The median calibration rate of samples by malignancy (where n ≥ 10 samples, excluding CTCL samples) was 88.1%, across a variety of fresh and archived sample sources (177 of 201 samples). TCR-β and/or TCR-γ clonotypes were identified in 40 of 95 samples (42.1%) from 66 patients with suspected CTCL. This NGS-MRD Assay is a valuable and sensitive tool for monitoring MRD in patients with plasma cell and lymphoid malignancies and assisting in the diagnosis of CTCL.

A clinician's guide to cutaneous T-cell lymphoma presenting as recalcitrant eczematous dermatitis in adults.

Cutaneous T-cell lymphoma (CTCL) encompasses a group of low-grade, non-Hodgkin lymphoma, including mycosis fungoides and Sézary syndrome. Diagnosis of CTCL can be challenging given the prolonged, gradual onset and shared characteristics with many benign inflammatory skin diseases. In this case series, we describe four unique cases of patients with chronic, recalcitrant eczematous dermatitis who presented for a patch-test consultation and were ultimately diagnosed with CTCL. In particular, we highlight clinical pearls to aid in distinguishing CTCL from inflammatory dermatoses and describe the diagnostic strategy to help dermatologists arrive at the diagnosis of CTCL at earlier stages of the disease.

Morphologic Features of Cutaneous T-Cell Lymphomas Using Dermoscopy and High Frequency Ultrasound.

The diagnosis of cutaneous T-cell lymphomas (CTCL) is frequently delayed by a median of three years and requires the clinical evaluation of an experienced dermatologist and a confirmatory skin biopsy. Dermoscopy and high-frequency ultrasound (HFUS) represent two non-invasive diagnostic tools. While dermoscopy is inexpensive and widely used for the diagnosis of melanoma and non-melanoma skin cancers, HFUS of skin lymphomas represents a novel diagnostic approach that is not yet implemented in the routine dermatologic practice. The aim of our study was to prospectively assess skin lesions of patients with either CTCL patches or plaques with dermoscopy and HFUS and to compare the findings with atopic dermatitis (AD) and psoriasis. Thirteen patients with an established diagnosis of CTCL, psoriasis, or AD were studied: Dermoscopy features including spermatozoa-like structures and the presence of white scales could assist in differentiating between early-stage CTCL and AD. HFUS measurements of the skin thickness indicated increased epidermal-, thickness in CTCL, and psoriasis compared with AD. Our results support the use of dermoscopy as a useful tool to diagnose CTCL. HFUS could augment the dermatologic assessment, but further studies will be needed to define standardized parameters.

Cutaneous T-cell lymphoma is associated with increased risk of lymphoma, melanoma, lung cancer, and bladder cancer

Patients with cutaneous T-cell lymphoma (CTCL) are at a higher risk of developing second malignancies. However, rates of incidence vary significantly across studies. A systematic review and meta-analysis of articles published between 1950 and 2019 was performed to evaluate the risk of second malignancies in patients with CTCL. We identified 10 eligible studies, including 12 patient cohorts, with 5.9% to 16.8% of patients developing second malignancies. All studies showed a male predominance for patients developing second malignancies. The mean age across the studies ranged from 44.6 to 68.0years. The time between the diagnosis of CTCL and second malignancy ranged from 2.1 to 5.4years (mean, 3.29y; 95% confidence interval [CI], 2.69-5.15). Meta-analysis showed a standardized incidence ratio of 2.18 (95% CI, 1.43-2.93) for all malignancies. The standardized incidence ratios were 15.25 (95% CI, 7.70-22.79) for Hodgkin lymphoma, 4.96 (95% CI, 3.58-6.33) for non-Hodgkin lymphoma, 1.69 (95% CI, 1.18-2.21) for lung cancer, 1.72 (95% CI, 1.18-2.21) for bladder cancer, and 3.09 (95% CI, 1.77-6.43) for melanoma. We find that patients with CTCL are at increased risk of second malignancies, especially Hodgkin and non-Hodgkin lymphoma, lung cancer, bladder cancer, and melanoma. These findings provide evidence of a population at increased risk of malignancy. Early detection may decrease the morbidity burden of second malignancies, thus providing a strong rationale for prospective screening studies.

110 RNA expression analysis in stage IVA-B cutaneous T-cell lymphoma to identify novel biomarkers of prognosis and diagnosis

Cutaneous T-cell lymphoma (CTCL) is a primary non-Hodgkin lymphoma and the most common primary cutaneous lymphoma. Due to its varied clinical and histological presentation, the accurate diagnosis of CTCL is difficult and therefore delayed; in advanced stages, CTCL is incurable and often fatal, though treatment-resistant phenotypes and survival time can be difficult to predict. There remains an unmet need for new and more effective diagnostic and prognostic indicators and novel therapies. To this end, malignant cells were isolated from the peripheral blood of 15 CTCL patients using antibody-magnetic bead or FACS sorting for CD3+CD4+ and CD7- and/or CD26- based on the known aberrant phenotype of the patient or positive selection for CD45RO+ CD4+ T cells in 3 healthy controls.

Sensitivity and specificity of T-cell receptor PCR BIOMED-2 clonality analysis for the diagnosis of cutaneous T-cell lymphoma.

Early and precise diagnosis of cutaneous T-cell lymphomas (CTCL) is challenging. Currently, polymerase chain reaction (PCR)-based clonality assessment of the T-cell receptor (TCR) is a helpful tool for this diagnosis. In this retrospective study, we aimed to assess the sensitivity and specificity of this method for the diagnosis of CTCL. Monoclonal rearrangement of the TCR was investigated retrospectively by PCR-based clonality assessment based on 292 DNA samples from skin biopsies of patients with a suspicion of CTCL. Algorithms were based on different ratios (three or five-fold difference) between the dominant PCR peak and the third highest PCR peak. A PCR peak five-fold higher than the third highest PCR peak demonstrated significantly greater specificity (83.7% versus 76.4%) but lower sensitivity (59.8% versus 68.6%) compared to a cut-off of three-fold higher. Our results confirm the diagnostic value of TCR clonality analysis which may be used to define the ideal cut-off in order to optimize sensitivity and specificity.

Expression of CCR3 and CCR4 Suggests a Poor Prognosis in Mycosis Fungoides and Sézary Syndrome.

Tumor cells in cutaneous T-cell lymphoma express limited numbers of chemokine receptors. We investigated the expression patterns of CXCR3, CCR3, CCR4 and CCR10 in mycosis fungoides, Sézary syndrome, lym-phomatoid papulosis and anaplastic large cell lymphoma in 121 skin biopsy samples. CXCR3 was expressed in 86% of mycosis fungoides cases but in no anaplastic large cell lymphoma cases. CCR3 was expressed in 73% of cases of CD30+ lymphoproliferative disorders such as lymphomatoid papulosis and anaplastic large cell lymphoma. Mycosis fungoides/Sézary syndrome patients with high CCR3 or CCR4 expression had a poorer survival prognosis than mycosis fungoides/Sézary syndrome patients whose tumor cells did not express these receptors. CCR10 was expressed in 50% of mycosis fungoides/Sézary syndrome cases and in 13% of cases with CD30+ lym-phoproliferative disorders. These results suggest that differential patterns of CXCR3, CCR3, CCR4 and CCR10 expression are useful for the diagnosis of cutaneous T-cell lymphoma. Moreover, expression of CCR3 or CCR4 suggests a poor prognosis in mycosis fungoides/Sézary syndrome.