Variant Of Cutaneous T-cell Lymphoma Research Articles

Sézary Syndrome as an Etiological Cause of Erythroderma: A Case Report

Exfoliative erythroderma is a syndrome characterized by erythema and scaling that affects more than 90% of the body surface. In contrast, Sézary Syndrome is a rare and harmful variant of cutaneous T-cell lymphoma, defined by a triad: circulation of neoplastic T-cells, erythroderma and lymphadenopathy, the latter of which may or may not be present. We report a case of an erythrodermic patient, with the etiological cause confirmed as Sézary Syndrome.

Cutaneous T cell Lymphoma in Skin of Color: A Review.

People of color (POC) affected by skin cancer suffer disproportionately from worse morbidity and mortality. Although skin cancers occur most frequently in White individuals overall, cutaneous T-cell lymphoma (CTCL) is an exception. CTCL is a rare skin cancer comprising several subtypes of non-Hodgkin lymphoma; each contains a unique clinical profile that varies with race. Our aim is to review and compile the differences in epidemiology, clinical presentation, treatments, and outcomes of the CTCL subtypes in Black, Asian or Pacific-Islander (API), and Hispanic patients. The current literature supports that there are nuances in the course of CTCL that differ with race. Across multiple studies, racial differences in incidence patterns have been reported, with the highest rates among Black patients. Cutaneous manifestation of CTCL are highly variable in POC, and the predilection for clinical CTCL variants often differs with race, as well as severity of cutaneous involvement (BSA). Response to and type of treatment also differs among POC, and may be partially attributable to the varying CTCL subtypes experienced by certain races. Prognostic factors tend to vary with race, although Black patients consistently experience poor outcomes, while API patients may have a more favorable prognosis. Currently, there is no definitive conclusion to account for differences observed in CTCL skin of color patients, however biologic and socioeconomic factors have been proposed as potential drivers. As POC comprise an increasing portion of our population, adequate physician awareness and knowledge of racial nuances in CTCL are necessary to begin addressing these disparities.

Staphylococcus aureus induces drug resistance in cancer T cells in Sézary syndrome

AbstractPatients with Sézary syndrome (SS), a leukemic variant of cutaneous T-cell lymphoma (CTCL), are prone to Staphylococcus aureus infections and have a poor prognosis due to treatment resistance. Here, we report that S aureus and staphylococcal enterotoxins (SE) induce drug resistance in malignant T cells against therapeutics commonly used in CTCL. Supernatant from patient-derived, SE-producing S aureus and recombinant SE significantly inhibit cell death induced by histone deacetylase (HDAC) inhibitor romidepsin in primary malignant T cells from patients with SS. Bacterial killing by engineered, bacteriophage-derived, S aureus–specific endolysin (XZ.700) abrogates the effect of S aureus supernatant. Similarly, mutations in major histocompatibility complex (MHC) class II binding sites of SE type A (SEA) and anti-SEA antibody block induction of resistance. Importantly, SE also triggers resistance to other HDAC inhibitors (vorinostat and resminostat) and chemotherapeutic drugs (doxorubicin and etoposide). Multimodal single-cell sequencing indicates T-cell receptor (TCR), NF-κB, and JAK/STAT signaling pathways (previously associated with drug resistance) as putative mediators of SE-induced drug resistance. In support, inhibition of TCR-signaling and Protein kinase C (upstream of NF-κB) counteracts SE-induced rescue from drug-induced cell death. Inversely, SE cannot rescue from cell death induced by the proteasome/NF-κB inhibitor bortezomib. Inhibition of JAK/STAT only blocks rescue in patients whose malignant T-cell survival is dependent on SE-induced cytokines, suggesting 2 distinct ways SE can induce drug resistance. In conclusion, we show that S aureus enterotoxins induce drug resistance in primary malignant T cells. These findings suggest that S aureus enterotoxins cause clinical treatment resistance in patients with SS, and antibacterial measures may improve the outcome of cancer-directed therapy in patients harboring S aureus.

MAb14, a Monoclonal Antibody against Cell Surface PCNA: A Potential Tool for Sezary Syndrome Diagnosis and Targeted Immunotherapy.

Mycosis fungoides (MF) and Sézary syndrome (SS) are the most common types of primary cutaneous T-cell lymphoma (CTCL). Proliferating cell nuclear antigen (PCNA) is expressed on the cell surface of cancer cells (csPCNA), but not on normal cells. It functions as an immune checkpoint ligand by interacting with natural killer (NK) cells through the NK inhibitory receptor NKp44, leading to the inhibition of NK cytotoxicity. A monoclonal antibody (mAb14) was established to detect csPCNA on cancer cells and block their interaction with NKp44. In this study, three CTCL cell lines and peripheral blood mononuclear cells (PBMCs) from patients with SS and healthy donors were analyzed for csPCNA using mAb14, compared to monoclonal antibody PC10, against nuclear PCNA (nPCNA). The following assays were used: immunostaining, imaging flow cytometry, flow cytometry, cell sorting, cell cycle analysis, ELISA, and the NK-cell cytotoxic assay. mAb14 successfully detected PCNA on the membrane and in the cytoplasm of viable CTCL cell lines associated with the G2/M phase. In the Sézary PBMCs, csPCNA was expressed on lymphoma cells that had an atypical morphology and not on normal cells. Furthermore, it was not expressed on PBMCs from healthy donors. In the co-culture of peripheral blood NK (pNK) cells with CTCL lines, mAb14 increased the secretion of IFN-γ, indicating the reactivation of pNK activity. However, mAb14 did not enhance the cytotoxic activity of pNK cells against CTCL cell lines. The unique expression of csPCNA detected by mAb14 suggests that csPCNA and mAb14 may serve as a potential biomarker and tool, respectively, for detecting malignant cells in SS and possibly other CTCL variants.

Spatially Guided and Single Cell Tools to Map the Microenvironment in Cutaneous T-Cell Lymphoma

Mycosis fungoides (MF) and Sézary syndrome (SS) are two closely related clinical variants of cutaneous T-cell lymphomas (CTCL). Previously demonstrated large patient-to-patient and intra-patient disease heterogeneity underpins the importance of personalized medicine in CTCL. Advanced stages of CTCL are characterized by dismal prognosis, and the early identification of patients who will progress remains a clinical unmet need. While the exact molecular events underlying disease progression are poorly resolved, the tumor microenvironment (TME) has emerged as an important driver. In particular, the Th1-to-Th2 shift in the immune response is now commonly identified across advanced-stage CTCL patients. Herein, we summarize the role of the TME in CTCL evolution and the latest studies in deciphering inter- and intra-patient heterogeneity. We introduce spatially resolved omics as a promising technology to advance immune-oncology efforts in CTCL. We propose the combined implementation of spatially guided and single-cell omics technologies in paired skin and blood samples. Such an approach will mediate in-depth profiling of phenotypic and molecular changes in reactive immune subpopulations and malignant T cells preceding the Th1-to-Th2 shift and reveal mechanisms underlying disease progression from skin-limited to systemic disease that collectively will lead to the discovery of novel biomarkers to improve patient prognostication and the design of personalized treatment strategies.

Healthcare Provider Experience in Diagnosing and Treating Cutaneous T-Cell Lymphoma.

Cutaneous T-cell lymphoma (CTCL) is a rare, heterogeneous group of non-Hodgkin lymphomas characterized by various clinical, molecular, and histopathologic features of the skin. Variants of CTCL share many clinical features with common inflammatory skin diseases such as atopic dermatitis and psoriasis, making accurate and early diagnosis challenging in clinical settings. Inappropriate treatment or a delay in diagnosis can lead to increased morbidity and mortality. Here, we report findings from an online survey that investigated dermatology community practice, knowledge, and education surrounding CTCL. An electronic survey of ten questions was developed and approved by physician experts in CTCL to assess experiences in diagnosing and treating CTCL among healthcare providers (HCPs). The survey was deployed to 10,600 US dermatology HCPs, including medical doctors (MDs), doctors of osteopathic medicine (DOs), nurse practitioners (NPs), and physician assistants (PAs) and excluding HCPs associated with CTCL centers of excellence. Among 44 HCPs who responded and were eligible for inclusion, 82% had diagnosed between one and ten CTCL cases in the last 5years. Most respondents (91%) reported that they include CTCL in their differential diagnoses after patients do not respond to treatment of more common conditions. Patients with CTCL were frequently diagnosed with other inflammatory dermatoses-most commonly dermatitis and psoriasis-before a CTCL diagnosis, and many were treated with ineffective therapies for years. The most common length of time before a CTCL diagnosis was made was between 1 and 3years, though 16% of HCPs reported that patients were treated for other diseases or skin conditions for ≥ 5years. Two-thirds of HCPs agreed that further education surrounding CTCL is needed. Given the infrequency of CTCL and its similar presentation to other common dermatologic conditions, increased education of CTCL is needed in the dermatology community to improve patient outcomes.

Phenotypic plasticity of malignant T cells in blood and skin of a Sézary syndrome patient revealed by single cell transcriptomics.

Sézary Syndrome (SS) is an aggressive leukemic variant of cutaneous T-cell lymphomas (CTCL). In SS patients, malignant T cells are circulating through the blood and cause erythroderma. To compare the transcriptome of single cells in blood and skin samples from a patient with advanced SS. We utilized combined single cell RNA and T-cell receptor (TCR) sequencing (scRNA-seq). We scrutinized the malignant T cells in blood and skin in an unbiased manner without pre-sorting of cells. We observed different phenotypes of the same monoclonal malignant T-cell population, confirmed by TCR sequencing and inferred copy number variation analysis. Malignant T cells present in the circulating blood expressed genes resembling central memory T cells such as CCR7, IL7R and CD27. In the skin, we detected two major malignant T-cell populations: One subpopulation was closely related to the malignant T cells from the blood, while the other subpopulation expressed genes reminiscent of skin resident effector memory T cells including GZMB and NKG7. Pseudotime analysis indicated crucial transcriptomic changes in the transition of malignant T cells between blood and skin. These changes included the differential regulation of TXNIP, a putative tumor suppressor in CTCL, and the adaptation to the hypoxic conditions in the skin. Tumor cell proliferation in the skin was supported by stimulating interactions between myeloid cells and malignant T cells. Using scRNA-seq we detected a high degree of functional heterogeneity within the malignant T-cell population in SS and highlighted crucial differences between SS cells in blood and skin.

TMEM244 Gene Expression as a Potential Blood Diagnostic Marker Distinguishing Sézary Syndrome from Mycosis Fungoides and Benign Erythroderma

Sézary syndrome (SS) is a rare, aggressive leukemic variant of cutaneous T-cell lymphoma (CTCL). The disease is characterized by severe erythroderma, generalized lymphadenopathy, and the presence of circulating Sézary cells that are a clonal proliferation of CD4+/CD45RO+ malignant T lymphocytes primarily involving the skin (Huet et al., 2006; Willemze et al., 2005). SS is a leukemic variant of CTCL and is closely related to mycosis fungoides (MF), a more indolent and slowly progressing type (Booken et al., 2008).

Single-cell RNA-sequencing reveals predictive features of response to pembrolizumab in Sézary syndrome

ABSTRACT The PD-1 inhibitor pembrolizumab is effective in treating Sézary syndrome, a leukemic variant of cutaneous T-cell lymphoma. Our purpose was to investigate the effects of pembrolizumab on healthy and malignant T cells in Sézary syndrome and to discover characteristics that predict pembrolizumab response. Samples were analyzed before and after 3 weeks of pembrolizumab treatment using single-cell RNA-sequencing of 118,961 peripheral blood T cells isolated from six Sézary syndrome patients. T-cell receptor clonotyping, bulk RNA-seq signatures, and whole-exome data were integrated to classify malignant T-cells and their underlying subclonal heterogeneity. We found that responses to pembrolizumab were associated with lower KIR3DL2 expression within Sézary T cells. Pembrolizumab modulated Sézary cell gene expression of T-cell activation associated genes. The CD8 effector populations included clonally expanded populations with a strong cytotoxic profile. Expansions of CD8 terminal effector and CD8 effector memory T-cell populations were observed in responding patients after treatment. We observed intrapatient Sézary cell heterogeneity including subclonal segregation of a coding mutation and copy number variation. Our study reveals differential effects of pembrolizumab in both malignant and healthy T cells. These data support further study of KIR3DL2 expression and CD8 immune populations as predictive biomarkers of pembrolizumab response in Sézary syndrome.

Combined High-Throughput Approaches Reveal the Signals Driven by Skin and Blood Environments and Define the Tumor Heterogeneity in Sézary Syndrome.

Simple SummarySézary syndrome (SS) is a leukemic and incurable variant of cutaneous T-cell lymphoma characterized by the accumulation of neoplastic CD4+ lymphocytes in the blood, lymph nodes, and skin. With the exception of allogenic transplantation, no curative chance is available to treat SS, and it is a priority to find new therapies that target SS cells within all disease compartments. This review aims to summarize the more recent analyses conducted on skin- and blood-derived SS cells concurrently obtained from the same SS patients. The results highlighted that skin-SS cells were more active/proliferating with respect to matched blood SS cells that instead appeared quiescent. These data shed the light on the possibility to treat blood and skin SS cells with different compounds, respectively. Moreover, this review recaps the more recent findings on the heterogeneity of circulating SS cells that presented a series of novel markers that could improve diagnosis, prognosis and therapy of this lymphoma.Sézary syndrome (SS) is an aggressive variant of cutaneous t-cell lymphoma characterized by the accumulation of neoplastic CD4+ lymphocytes—the SS cells—mainly in blood, lymph nodes, and skin. The tumor spread pattern of SS makes this lymphoma a unique model of disease that allows a concurrent blood and skin sampling for analysis. This review summarizes the recent studies highlighting the transcriptional programs triggered by the crosstalk between SS cells and blood–skin microenvironments. Emerging data proved that skin-derived SS cells show consistently higher activation/proliferation rates, mainly driven by T-cell receptor signaling with respect to matched blood SS cells that instead appear quiescent. Biochemical analyses also demonstrated an hyperactivation of PI3K/AKT/mTOR, a targetable pathway by multiple inhibitors currently in clinical trials, in skin SS cells compared with a paired blood counterpart. These results indicated that active and quiescent SS cells coexist in this lymphoma, and that they could be respectively treated with different therapeutics. Finally, this review underlines the more recent discoveries into the heterogeneity of circulating SS cells, highlighting a series of novel markers that could improve the diagnosis and that represent novel therapeutic targets (GPR15, PTPN13, KLRB1, and ITGB1) as well as new genetic markers (PD-1 and CD39) able to stratify SS patients for disease aggressiveness.

Genetically Driven CD39 Expression Affects Sezary Cell Viability and IL-2 Production and Detects Two Patient Subsets with Distinct Prognosis

Sézary syndrome (SS) is a rare and aggressive variant of cutaneous T-cell lymphoma. It is characterized by the copresence of CD4+ neoplastic lymphocytes, named Sezary cells, mainly in the blood, lymph nodes, and skin where they induce chronic inflammation that in turn impairs the patient's QOL and fuels neoplastic cells. SS is not readily cured, but immunotherapy is becoming an effective option for this lymphoma. In this study, we investigated, in a large cohort of patients with SS, the expression and function of the immune checkpoint molecule CD39, which degrades proinflammatory extracellular adenosine triphosphate. We showed that the SNP rs10748643 A/G within the ENTPD1 gene coding for the CD39 protein controls its expression level. Patients carrying the A/G‒G/G genotype showed a significantly higher frequency of clonal CD4+CD39+ SS cells than those carrying the A/A genotype. Different from other cancers, high CD39 expression correlates with a better prognosis. Comparing primary G/G with A/A lymphoma cells, we observed that G/G SS cells have a higher ability to degrade adenosine triphosphate, increased apoptotic susceptibility, and upon activation, reduced IL-2 production. Accordingly, CD39 enzymatic inhibition enhances SS cell viability and IL-2 production on activation. These results strongly suggest a special caution for SS treatment with therapeutic inhibitors of CD39.

Understanding Cell Lines, Patient-Derived Xenograft and Genetically Engineered Mouse Models Used to Study Cutaneous T-Cell Lymphoma.

Cutaneous T cell lymphoma (CTCL) is a spectrum of lymphoproliferative disorders caused by the infiltration of malignant T cells into the skin. The most common variants of CTCL include mycosis fungoides (MF), Sézary syndrome (SS) and CD30+ Lymphoproliferative disorders (CD30+ LPDs). CD30+ LPDs include primary cutaneous anaplastic large cell lymphoma (pcALCL), lymphomatoid papulosis (LyP) and borderline CD30+ LPD. The frequency of MF, SS and CD30+ LPDs is ~40–50%, <5% and ~10–25%, respectively. Despite recent advances, CTCL remains challenging to diagnose. The mechanism of CTCL carcinogenesis still remains to be fully elucidated. Hence, experiments in patient-derived cell lines and xenografts/genetically engineered mouse models (GEMMs) are critical to advance our understanding of disease pathogenesis. To enable this, understanding the intricacies and limitations of each individual model system is highly important. Presently, 11 immortalized patient-derived cell lines and different xenograft/GEMMs are being used to study the pathogenesis of CTCL and evaluate the therapeutic efficacy of various treatment modalities prior to clinical trials. Gene expression studies, and the karyotyping analyses of cell lines demonstrated that the molecular profile of SeAx, Sez4, SZ4, H9 and Hut78 is consistent with SS origin; MyLa and HH resemble the molecular profile of advanced MF, while Mac2A and PB2B represent CD30+ LPDs. Molecular analysis of the other two frequently used Human T-Cell Lymphotropic Virus-1 (HTLV-1)+ cell lines, MJ and Hut102, were found to have characteristics of Adult T-cell Leukemia/Lymphoma (ATLL). Studies in mouse models demonstrated that xenograft tumors could be grown using MyLa, HH, H9, Hut78, PB2B and SZ4 cells in NSG (NOD Scid gamma mouse) mice, while several additional experimental GEMMs were established to study the pathogenesis, effect of drugs and inflammatory cytokines in CTCL. The current review summarizes cell lines and xenograft/GEMMs used to study and understand the etiology and heterogeneity of CTCL.

The Role of Tumor Microenvironment in the Pathogenesis of Sézary Syndrome.

Sézary syndrome is an aggressive leukemic variant of cutaneous T-cell lymphomas, characterized by erythroderma, lymphadenopathy, and peripheral blood involvement by CD4+ malignant T-cells. The pathogenesis of Sézary syndrome is not fully understood. However, the course of the disease is strongly influenced by the tumor microenvironment, which is altered by a combination of cytokines, chemokines, and growth factors. The crosstalk between malignant and reactive cells affects the immunologic response against tumor cells causing immune dysregulation. This review focuses on the interaction of malignant Sézary cells and the tumor microenvironment.

A Fatal Case of Febrile Ulceronecrotic Mucha-Habermann Disease in a 10-Year-Old Boy.

A Fatal Case of Febrile Ulceronecrotic Mucha-Habermann Disease in a 10-Year-Old Boy.

Sezary Syndrome: A Clinico-biological Study of 5 Cases

<i>Background</i>: Sézary syndrome (SS) is a rare erythrodermic and leukemic variant of cutaneous T-cell lymphoma (CTCL) that belongs to the group of non-Hodgkin's lymphomas (NHL) resulting from malignant proliferation of skin-homing T cells. We report through a series of 5 cases, the experience of the hematology laboratory in the diagnosis of the syndrome of Sézary. <i>Methods</i>: This is a retrospective study of 5 cases of Sezary syndrome collected in the dermatology department of Marrakech. <i>Results</i>: five patients were identified with the clinicopathological criteria of SS. At the time of diagnosis, all 5 patients had erythroderma and generalized lymphadenopathy in both superficial and deep stations. The white blood cell count was elevated (>10,000 WBC/mm³) in all 5 patients with a mean value of 18,120 WBC/mm³. The blood smear showed the presence of 75% (27 G/l) of small to medium-sized cells with a high nucleocytoplasmic ratio and cerebriform nuclei typical of Sezary cells and suggests the diagnosis of SS. <i>Conclusions</i>: The diagnosis of SS remains a challenge in many situations, the pathophysiology and definition of SS have evolved significantly over the past decades.

Granulomatous Slack Skin with Hypercalcemia Indolently Mimicking Sarcoidosis

&lt;p class="abstract"&gt;Granulomatous slack skin is a rare variant of cutaneous T-cell lymphoma characterized by lax skin and granulomatous infiltrate with loss of elastic fibers on histology. We report a unique case of a female presenting with CD30-granulomatous slack skin complicated by hypercalcemia, initially diagnosed, and managed as sarcoidosis. Interestingly, she had a history of previously treated CD30+ cutaneous T-cell lymphoma. Granulomatous slack skin can frequently mimic other benign and malignant cutaneous diseases, prompting the need for clinical vigilance from dermatologists.&lt;/p&gt;

Epidemiology and status of specialized medical care for patients with cutaneous T-cell lymphomas: A survey of 163 patients of Russian Federation

Background. In the Russian Federation, there are currently no official statistical data concerning the incidence of cutaneous T-cell lymphomas (CTCL) and the prevalence of CTCL due to the absence of isolation rubrics in the headings of lymphoproliferative diseases of the federal state statistical observation of the separate neologies.&#x0D; Aim. To obtain and analyze the results of the number of patients with CTCL (mycosis fungoides and Szary syndrome) who were on the dispensary observation in medical organizations of dermatovenereological profile in the period 2015 to 2020, the demographic and clinical-epidemiological characteristics, the applied methods of diagnosis and the therapy.&#x0D; Materials and methods. To conduct the study, the questionnaire was developed to obtain the information of the number of patients observed with diagnoses of "mycosis fungoides" and "Szary syndrome" in medical organizations of the dermatovenereological profile of the Federal Subjects of the Russian Federation in the period 2015-2020, were researched the demographic characteristics, the diagnostic methods and the therapy in the studied group of patients.&#x0D; Results. From 24 Federal Subjects of the Russian Federation were obtained the data concerning 163 patients with CTCL under the observation in medical organizations in the period 20152020. From 35 Federal Subjects of the Russian Federation the information showed the absence of patients with CTCL under the observation in medical organizations of dermatovenereological profile. We did not receive the data from 10 Federal Subjects of the Russian Federation. Among 163 patients in 144 (89%) were diagnosed mycosis fungoides, in 17 (10%) Szary syndrome and in 2 (1%) patients were diagnosed other variants of CTCL. The data concerning the demographic characteristics, the diagnosis and the therapy of CTCL were obtained in 155 of 167 patients. We showed that in 44% of cases, the disease was detected by dermatovenerologists, and the diagnosis was most often (39%) determined by oncologists. In 49% of cases the diagnosis was determined only on the basis of the results of histological study, the immunohistochemical study and the polymerase chain reaction were used in 33% and in 3% of cases, respectively. From 155 patients, 52% were under dispensary observation by oncologists, 41% by hematologists, 26% of patients were observed by dermatologists; 99 patients were under dispensary observation by only one specialist: 50 (32%) of patients by oncologist, 40 (26%) of patients by hematologist, 9 (6%) of patients by dermatovenerologist. Nine (6%) of patients were under dispensary observation by three specialists. The information concerning the therapy was available in 92 (59%) of 155 patients. The most commonly applied method of treatment was chemotherapy 72%, phototherapy was received by 26%, the same percentage (18%) was received in two groups: the application of interferon 2b and the application of methotrexate, 1% of patients received radiation therapy. The lethal outcome was registered in 53 (34%) patients. Median duration of the disease from the time of diagnosis to death was 3.55.0 years, the median 2 years, the mode 1 year.&#x0D; Conclusion. For the first time, the attempt was made to summarize the data of the number of patients with CTCL, to describe the demographic, clinical and epidemiological characteristics, the data concerning diagnostics and the therapy. The obtained preliminary results are required further detailing investigation in close cooperation with the professional specialists such as hematologists and oncologists.

Woringer-Kolopp Disease, a Rare Dermatologic Malignancy Presenting in a Young Male

Abstract Pagetoid reticulosis (PR), also known as Woringer-Kolopp disease (WKD), is a rare variant of cutaneous T-cell lymphoma usually presenting as an isolated patch or plaque on the extremities. Immunohistochemical staining of T-cell markers can be variable, but as the name implies the epidermotropic lymphocytes consistently display a “pagetoid” appearance. Two variants of the disease are described: the localized type WKD and the disseminated type (Ketron-Goodman disease or KGD). We present the case of a 31 years old patient with no other known personal history, who is a dentist working with irritating chemicals such as orthophosphoric acid 36% and hydrofluoric acid 9.5%, who presented to our clinic for a rash consisting of papulo-vesicles with a diameter of 0,5-1.3 cm, intensely pruritic, firm to palpation, arranged on a diffuse erythematous background, located on the left hand on metacarpophalangeal (MCP) joints I-II, that had been evolving for about 1 year, with healing under local cortisone treatment but rapid relapse with local extension of lesions. Diagnosis is confirmed via a skin biopsy of the affected area followed by a cytologic examination. Histopathological appearance and immunophenotypic profile of the lymphoid infiltrate (CD3+, CD4-, CD8+) supported the diagnosis of PR, the localized form WKD. Treatment typically entails topical corticosteroids, alkylating agents, and retinoids but as an additional therapy in cases with localized form, narrowband ultraviolet B (NB-UVB) with remission of lesions and their recurrence, has also been chosen for our pacient.

Targeting Epigenetic Modifiers Can Reduce the Clonogenic Capacities of Sézary Cells.

Sézary syndrome (SS) is an aggressive leukemic variant of cutaneous T-cell lymphomas (CTCL) in which the human Telomerase Reverse Transcriptase (hTERT) gene is re-expressed. Current available treatments do not provide long-term response. We previously reported that Histone deacetylase inhibitors (HDACi, romidespin and vorinostat) and a DNA methyltransferase inhibitor (DNMTi, 5-azacytidine) can reduce hTERT expression without altering the methylation level of hTERT promoter. Romidepsin and vorinostat are approved for CTCL treatment, while 5-azacytidine is approved for the treatment of several hematological disorders, but not for CTCL. Here, using the soft agar assay, we analyzed the functional effect of the aforementioned epidrugs on the clonogenic capacities of Sézary cells. Our data revealed that, besides hTERT downregulation, epidrugs’ pressure reduced the proliferative and the tumor formation capacities in Sézary cells in vitro.

Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from S&amp;#233;zary Syndrome Patients for Transcriptomic Profiling

Cutaneous T-cell lymphomas (CTCL) are derived from the transformation and uncontrolled proliferation of mature skin-homing T cells, and mycosis fungoides (MF) and Sézary syndrome (SS) represent the most common subtypes. Despite a number of studies on characterizing gene expression, genetic alterations, and epigenetic abnormalities of CTCL, the molecular pathogenesis of MF/SS remains unclear. MF refers to the more common CTCL with a skin-predominance, and is usually limited to skin, whereas SS is an aggressive leukemic variant of CTCL with widespread skin involvement and is characterized by neoplastic distribution mainly involving blood, skin, and lymph node. Focusing on clinical practice, the identification of gene expression biomarkers has enormous potential to improve diagnosis and treatment of MF/SS. Indeed, recent transcriptomic studies have identified potential diagnostic biomarkers from differences in gene expression between normal and malignant T cells, which may improve our understanding of SS biology, and reveal potential therapeutic targets. This manuscript describes a detailed reproducible protocol for the isolation of peripheral blood mononuclear cells from fresh whole blood from patients diagnosed with SS, selection of CD4+ memory T cells (CD4+CD45RO+ T cells), chemical stimulation, and preparation of RNA suitable for transcriptomic profiling to discover novel prognostic molecular markers to gain additional insight in disease etiology. The stimulation using chemical agonist to activate nuclear regulation provides more specific assessment for pathways important in the dynamic transcription regulation and gene expression and eliminates confounding defects that may arise from upstream signaling defects arising from TCR antigen loss at the cell membrane. The data obtained from comparison of transcriptome of unstimulated to stimulated SS T cells unmasks functional regulatory gene expression defects not evident from analysis of quiescent unstimulated cells. Furthermore, the method outlined from this approach can be adapted for studying T cell gene expression defects in other T cell immune diseases.