Aggressive T-cell Lymphoma Research Articles

Characterization of a novel canine T-cell line established from a spontaneously occurring aggressive T-cell lymphoma with large granular cell morphology

Dogs with lymphoma are established as good model for human non-Hodgkin lymphoma studies. Canine cell lines derived from lymphomas may be valuable tools for testing new therapeutic drugs. In this context, we established a canine T-cell line, PER-VAS, from a primary aggressive T-cell lymphoma with large granular morphology. Flow cytometric analysis revealed a stable immunophenotype: PER-VAS cells were positively labelled for CD5, CD45, MHC II and TLR3, and were negative for CD3, CD4 and CD8 expression. Although unstable along the culture process, IL-17 and MMP12 proteins were detectable as late as at passages 280 and 325i.e. respectively 24 and 29 months post isolation. At passage 325, PER-VAS cells maintained the expression of IL-17, CD3, CD56, IFNγ and TNFα mRNAs as shown by RT-PCR analysis. Stable rearrangement of the TCRγ gene has been evidenced by PCR. PER-VAS cells have a high proliferation index with a doubling time of 16.5h and were tumorigenic in Nude mice. Compared to the canine cell lines already reported, PER-VAS cells display an original expression pattern, close to NKT cells, which makes them valuable tools for in vitro comparative research on lymphomas.

Anti-thymocyte globulin could improve the outcome of allogeneic hematopoietic stem cell transplantation in patients with highly aggressive T-cell tumors

The early experiment result in our hospital showed that anti-thymocyte globulin (ATG) inhibited the proliferation of lymphoid tumor cells in the T-cell tumors. We used the ATG as the part of the conditioning regimen and to evaluate the long-term anti-leukemia effect, the safety and complication in the patients with highly aggressive T-cell lymphomas. Twenty-three patients were enrolled into this study. At the time of transplant, six patients reached first or subsequent complete response, three patients had a partial remission and 14 patients had relapsed or primary refractory disease. The conditioning regimen consisted of ATG, total body irradiation, toposide and cyclophosphamide. The complete remission rate after transplant was 95.7%. At a median follow-up time of 25 months, 16 (69.6%) patients are alive and free from diseases, including nine patients in refractory and progressive disease. Seven patients died after transplant, five from relapse and two from treatment-related complications. The incidence of grades II–IV acute graft-vs-host disease (GvHD) was 39.1%. The maximum cumulative incidence of chronic GvHD was 30%. The most frequent and severe conditioning-related toxicities observed in 8 out of 23 patients were grades III/IV infections during cytopenia. Thus, ATG-based conditioning is a feasible and effective alternative for patients with highly aggressive T-cell tumors.

The aggressive peripheral T-cell lymphomas: 2015.

T-cell lymphomas make up approximately 10%-15% of lymphoid malignancies. The frequency of these lymphomas varies geographically, with the highest incidence in parts of Asia. The diagnosis of aggressive peripheral T-cell lymphoma (PTCL) is usually made using the World Health Organization classification. The ability of hematopathologists to reproducibly diagnosis aggressive PTCL is lower than that for aggressive B-cell lymphomas, with a range of 72%-97% for the aggressive PTCLs. Patients with aggressive PTCL are staged using the Ann Arbor Classification. Although somewhat controversial, positron emission tomography scans seem to be useful as they are in aggressive B-cell lymphomas. The most commonly used prognostic index is the International Prognostic Index. The specific subtype of aggressive PTCL is an important risk factor, with the best survival seen in anaplastic large-cell lymphoma-particularly young patients with the anaplastic lymphoma kinase positive subtype. Anaplastic large-cell lymphoma is the only subgroup to have a good response to a CHOP-like regimen. Angioimmunoblastic T-cell lymphoma has a prolonged disease-free survival in only ~20% of patients, but younger patients who have an autotransplant in remission seem to do better. PTCL-not otherwise specified is not one disease. Anthracycline-containing regimens have disappointing results, and a new approach is needed. Natural killer/T-cell lymphoma localized to the nose and nasal sinuses seems to be best treated with radiotherapy-containing regimens. Enteropathy-associated PTCL and hepatosplenic PTCL are rare disorders with a generally poor response to therapy, although selected patients with enteropathy-associated PTCL seem to benefit from intensive therapy.

Clinical analysis of 25 patients with aggressive peripheral T-cell lymphoma in advanced stage treated with autologous stem cell transplantation

目的探讨大剂量化疗联合自体造血干细胞移植（ASCT）治疗侵袭性外周T细胞淋巴瘤（PTCL）的疗效。方法回顾性分析1997年5月至2013年6月于完全缓解（CR）期接受ASCT治疗的25例侵袭性PTCL患者的临床资料。结果①25例患者中男16例，女9例，中位发病年龄30(12~54）岁。病理类型包括PTCL非特指型（PTCL-U) 16例、血管免疫母细胞淋巴瘤（AITL）4例、间变大细胞淋巴瘤（ALCL）3例和肝脾T细胞淋巴瘤（HSTL) 2例。临床分期Ⅲ期和Ⅳ期分别为8例和17例；其中合并骨髓侵犯者9例。移植前19例处于CR1期，另6例为CR2期。②2例HSTL均为骨髓侵犯、国际预后指数评分高危患者，联合化疗后获得CR1并行ASCT巩固治疗，但均于移植后12个月内早期复发、死亡。其余23例患者中位随访38(14~110）个月，3年无进展生存（PFS）率和总体生存（OS）率分别为（63.1±10.5）%和（71.8±9.9）%。CR1期（17例）移植者的生存有优于CR2期（6例）移植者的趋势[3年PFS率（74.9±11.0）％对（33.3±19.2）%，P=0.092; 3年OS率（80.2±10.4）％对（50.0±20.4）%，P=0.043]。无骨髓侵犯者的生存有优于骨髓侵犯者的趋势[3年PFS率（77.9±11.3）%对（40.0±17.4）%，P=0.142；OS率（84.4±10.2）％对（53.3±17.3）%，P=0.076]。结论ASCT改善化疗敏感侵袭性PTCL患者的生存。移植前非CR1状态和淋巴瘤骨髓侵犯可能是影响ASCT疗效的不良预后因素。HSTL患者预后差，ASCT后早期复发率高，需要进一步探索有效的治疗方案。

Hematopoietic stem cell transplantation for primary cutaneous γδ T-cell lymphoma and refractory subcutaneous panniculitis-like T-cell lymphoma

The panniculitic T-cell lymphomas (TCLs) comprise 2 distinct entities, αβ subcutaneous panniculitis-like TCL (SPTCL) and the γδ cutaneous TCLs with pannicular involvement primary cutaneous γδ (PCGD)-TCL. Although outcomes for most patients with SPTCL are favorable, those with PCGD-TCLs generally have an inferior outcome, and treatment strategies have not been well defined. Allogeneic hematopoietic stem cell transplantation (HSCT) has been shown to be a potentially curative strategy in aggressive TCLs and in refractory and advanced-stage mycosis fungoides. We sought to analyze the outcomes of HSCT for panniculitic cutaneous TCL. Fourteen patients (4 SPTCL, 10 PCGD-TCL) presented with primarily pannicular T-cell infiltrates. Seven patients underwent allogeneic HSCT from matched-related donors and matched-unrelated donors of which 4 (57%) are alive (1 SPTCL, 3 PCGD-TCL) at 7.8, 6.9, 6.2, and 0.25 years. Two patients underwent autologous HSCT (1 SPTCL, 1 PCGD-TCL) and both are alive at a median follow-up of 1.91 years. This study is limited by its retrospective nature and small sample size because of the rarity of SPTCL and PCGD-TCL. Aggressive therapy followed by allogeneic HSCT is a promising treatment modality for patients with PCGD-TCL.

Expression of T-follicular helper markers in sequential biopsies of progressive mycosis fungoides and other primary cutaneous T-cell lymphomas.

T-follicular helper (Tfh) lymphocytes represent the neoplastic cells of angioimmunoblastic T-cell lymphoma and have been observed also in several cutaneous T-cell lymphomas (CTCLs) and extracutaneous T-cell lymphomas, including peripheral T-cell lymphoma, not otherwise specified, mycosis fungoides (MF), cutaneous CD4 small/medium T-cell lymphoma (CD4SMTCL), and Sezary syndrome. We studied a large number of different types of primary CTCL for expression of Tfh markers, including 36 biopsies from 21 patients with MF (with sequential biopsies from patch stage and tumor stage of 15 patients), 13 patients with CD4SMTCL, 9 with lymphomatoid papulosis, 11 with cutaneous anaplastic large cell lymphoma (cALCL), 2 with cutaneous γ/δ T-cell lymphoma, 8 with subcutaneous panniculitis-like T-cell lymphoma, 3 with cutaneous aggressive epidermotropic CD8 cytotoxic T-cell lymphoma, 6 with cutaneous peripheral T-cell lymphoma, not otherwise specified, and 1 with Sezary syndrome. Expression of at least 3 of 5 markers (PD-1, CXCL-13, ICOS, Bcl-6, and CD10) in >10% of tumor cells was observed in 33 biopsies (MF = 20; CD4SMTCL = 11; 1 each cutaneous anaplastic large cell lymphoma and subcutaneous panniculitis-like T-cell lymphoma, respectively). Our study shows that a Tfh phenotype is very common in MF and CD4SMTCL but can be observed rarely also in other types of CTCL. Expression of Tfh markers should not be used for classification of any entity of CTCL and may only integrate other immunohistochemical stainings for a more accurate characterization of these disorders. Precise distinction of Tfh-positive CTCLs from secondary skin manifestations of angioimmunoblastic T-cell lymphoma cannot rest on demonstration of a Tfh phenotype alone and should be achieved by a synthesis of clinical, histological, and phenotypic features.

Cutaneous lymphomas: new entities and rare variants

Primary cutaneous lymphomas are the second most common group of extranodal non-Hodgkin lymphomas. Recently several new variants and entities have been described but have not yet become part of the World Health Organization (WHO) classification. These forms include the granulomatous form of mycosis fungoides, which is associated with a poorer prognosis, as well as indolent CD8+ lymphoproliferations on the head and at acral localizations. Within the group of cutaneous CD30+ lymphoproliferative disorders, new histological types of lymphomatoid papulosis have been identified, such as type D (CD8+ epidermotropic) and typeE (angioinvasive) which simulate aggressive lymphomas. Cutaneous peripheral T-cell lymphomas are a prognostically heterogeneous group of cutaneous lymphomas. The cutaneous CD8+ aggressive epidermotropic cytotoxic T-cell lymphoma and cutaneous gamma/delta T-cell lymphoma are very aggressive neoplasms, whereas cutaneous CD4+ small to medium-sized T-cell lymphoma in its solitary or localized form represents an indolent lymphoproliferation: the terminology, histogenesis and differentiation from nodular T-cell pseudolymphoma are still a matter of debate. Among B-cell lymphomas, disorders associated with Epstein-Barr virus (EBV) are discussed focusing on EBV diffuse large B-cell lymphoma of the elderly and EBV-associated mucocutaneous ulcer. This review describes the clinical, histological and immunophenotypic features of new and rare entities and variants of cutaneous lymphomas and highlights the impact of the clinicopathological correlation in the diagnostic process.

Composite lymphoma: Simultaneous development of three different lymphomas in a single patient: A clinical case

In the past decades, there has been an increase in the incidence of malignancies, including polyneoplasms. Composite synchronous lymphoma characterized by the simultaneous development of several lymphoproliferative diseases occurs very rarely. This paper describes a case of development of 3 different lymphomas in the same patient. Aggressive T-cell lymphoma and indolent B-cell lymphoma are diagnosed during lifetime; primary central nervous system lymphoma is verified only by examination of autopsy material. Three different lymphomas could be identified by an integrated assessment of their clinical picture and by current diagnostic techniques, such as histology, immunohistochemistry, and molecular genetics.

γδ T-cell-rich variants of pityriasis lichenoides and lymphomatoid papulosis: benign cutaneous disorders to be distinguished from aggressive cutaneous γδ T-cell lymphomas.

T cells with a γδ phenotype have been associated with aggressive lymphomas. Yet, inflammatory skin disorders and low-grade lymphoproliferative disorders have rarely been described with a predominant γδ T-cell infiltrate. To review our experience and determine the clinical relevance of the γδ T-cell phenotype in lymphomatoid papulosis (LyP) and pityriasis lichenoides (PL). A retrospective dermatopathology file review looking for LyP and PL characterized by a γδ T-cell phenotype was performed. Clinical manifestations and course, histological features and molecular data were analyzed. Six of 16 cases of LyP and four of 23 cases diagnosed as PL during a 5-year period (2009-14) were identified. The median follow-up for the whole group was 16months (range 3-64), showing an indolent clinical course in all cases. The detection of a predominantly γδ T-cell phenotype in papular lymphoid-rich infiltrates in the absence of other lesions is not associated with a clinically aggressive course. γδ T-cell-rich variants of LyP and PL may reflect a spectrum of related conditions. This is a single academic centre retrospective chart review of a relatively small sample.

Clinical Limitations of Surveillance Fluorine 18 Fluorodeoxyglucose Positron Emission Tomography-Computed Tomography According to Histologic Subtypes in Patients with Malignant Lymphoma Who Achieved Complete Remission

Abstract Introduction Fluorine 18 Fluorodeoxyglucose (FDG) Positron emission tomography-computed tomography (PET-CT) scan was highly sensitive modality for evaluating the staging or response of malignant lymphoma. The role of PET to detect early relapse in complete remission was still uncertain. We evaluated the clinical implication of surveillance PET-CT scan in lymphoma patients according to histologic subtype of lymphoma. Patients and methods A total 1009 patients of 1553 treated lymphoma patients, who achieved complete response following primary treatment, were retrospectively reviewed. Among these patients, we analyzed 202 patients who showed positive results of PET-CT and performed tissue biopsy of FDG uptake sites. Results Total 202 surveillance PET-CT positive patients, 120 (59.4%) patients were confirmed lymphoma relapse, 82 (40.6%) patients were histologically negative. Histologically negative results of 82 patients as follows: most (59.7%) patients were inflammation, 10 (2.1%) patients were solid tumor and 3 patients were tuberculosis. According to histologic subtype, PPV of Hodgkin's lymphoma was 25.0% (4/16), diffuse large B-cell lymphoma patients was 51.0% (47/92), mantle cell lymphoma (MCL) was 86.6% (13/15) and extranodal NK/T cell lymphoma (ENKL) was 85.7% (12/14) (Table 1). There was significant difference between PPV of aggressive B-cell non-Hodgkin lymphoma (NHL) except MCL and aggressive T-cell NHL (51.0% vs 78.5%, P=0.040). Indolent NHL was related with high PPV compared to aggressive B-cell NHL (70.2% vs 51.0%, P=0.032). Six (12.7%) patients of 47 indolent lymphoma were transformed to DLBCL at relapse. Four (8.5%) patients of 47 DLBCL patients showed histologic change to indolent lymphoma. There was no difference of predictive value according to stage, the number of extranodal involvement, lactic dehydrogenase and international prognostic index. PPV of FDG uptake in lymph node was 58.3% (63/108) and other sites except lymph node were 60.6% (57/94). Biopsy of stomach was 53.3% (8/15), small or large bowel was 55.5% (7/12). True positive patients of core biopsy was 61 (65.5%) of 93 patients, operation or excisional biopsy was 59 (67.0%) of 88 patients and aspiration cytology was 4 (19.0%) of 21 patients. Conclusion PPV of surveillance PET-CT in aggressive B-cell lymphoma was lower than indolent lymphoma or aggressive T-cell lymphoma. Surveillance PET-CT had clinical limitations according to histologic subtype of lymphoma. Histologic confirm by re-biospy should be recommended to exclude the false positive in aggressive B-cell lymphoma and distinguish the histologic transformation in indolent lymphoma. Table 1. Positive predictive value according to lymphoma subtype Pathology Lymphoma subtype positive negative Positive predictive value DLBCL (n=92) 47 45 51.0 MZBCL (n=26) 17 9 65.3 Follicular lymphoma (n=20) 16 4 80.0 Mantle cell lymphoma (n=15) 13 2 86.6 Burkitt lymphoma (n=3) 1 2 33.3 Plasmablastic lymphoma (n=1) 1 0 100 Lymphoplasmacytic lymphoma (n=1) 0 1 0 ENKL (n=14) 12 2 85.7 PTCL (n=11) 7 4 63.6 Mycosis fungoides (n=3) 2 1 66.6 Hodgkin's lymphoma (n=16) 4 12 25.0 Total (n=202) 120 82 59.4 Disclosures No relevant conflicts of interest to declare.

Simultaneous occurrence of a CD30 positive/ALK-negative high grade T-cell lymphoma and plasma cell myeloma: Report of a case

Simultaneous occurrences of T-cell and B-cell neoplasms are rare, and etiological relationships between these two malignancies are poorly understood. We report the case of a 76-year-old man who presented with hypercalcemia, multiple skin nodular lesions, fatigue, episodic fever, and night sweats. PET/CT scan showed diffuse skin and subcutaneous fat plane active lesions, supra- and infra- diaphragmatic active lymph nodes, liver and spleen involvement, bone marrow infiltration, and nonspecific bilateral lung nodules. A skin biopsy showed a high grade CD30-positive/ALK-negative T-cell lymphoma. A bone marrow biopsy showed involvement by the same neoplastic cells. Additionally, a monoclonal lambda restricted plasma cell population (15% of marrow elements) was identified, which, in view of an IgA lambda spike in the serum, was consistent with plasma cell myeloma. To the best of our knowledge, this case is the first reported case of a plasma cell neoplasm associated with an aggressive CD30-positive ALK-negative systemic T-cell lymphoma with skin involvement. Reporting such cases is important as it adds to the pool of rare cases of concomitant T-cell neoplasms and plasma cell myelomas, and might help in determining an etiological relationship, if any, between these two hematological malignancies.

Mogamulizumab and the treatment of CCR4-positive T-cell lymphomas.

Glyco-engineering has been developed to enhance the pharmacological properties of monoclonal antibodies (mAbs) resulting in superior immune effector function. Mogamulizumab is the first approved glyco-engineered therapeutic antibody and first approved mAb to target the CC chemokine receptor 4 (CCR4). CCR4 is principally expressed on Tregs and helper T cells (Th) where it functions to induce homing of these leukocytes to sites of inflammation. Tregs play an essential role in maintaining immune balance; however, in malignancy, Tregs impair host antitumor immunity and provide a favorable environment for tumors to grow. CCR4 is highly expressed by aggressive peripheral T-cell lymphomas (PTCLs), particularly adult T-cell leukemia/lymphoma (ATL) and cutaneous T-cell lymphomas (CTCLs). Mogamulizumab is a humanized anti-CCR4 mAb with a defucosylated Fc region that enhances antibody-dependent cellular cytotoxicity (ADCC). In addition, mogamulizumab depletes CCR4(+) Tregs, potentially evoking antitumor immune responses by autologous effector cells. This ability is highly pertinent as subsets of malignant T cells are believed to function as CD4(+) Tregs, overexpressing CCR4. Clinical trials with mogamulizumab have demonstrated clinical efficacy and tolerability for the treatment of relapsed/refractory aggressive T-cell lymphomas, previously associated with very poor outcomes.

Type D Lymphomatoid Papulosis: An uncommon Variant. A case report and review of the literature

Lymphomatoid papulosis (LyP) is an indolent form of primary cutaneous T-cell lymphoma, currently classified together with primary cutaneous anaplastic large T-cell lymphoma within the spectrum of CD30-positive lymphoproliferative disorders. It is characterized by presenting as a clinically benign but histopathological malignant disease. Clinical features consist in recurrent waxing and waning red papules. Histopathologically, there are 4 variants recognized, Type A or Hystiocitic type, being the most frecuent of all, Type B or Mycosis fungoides-like, Type C or Anaplastic large-cell lymphoma-like and Type D, the most recently described and uncommon variant with features similar to Cutaneous Aggressive CD8-Positive Cytotoxic T-Cell Lymphoma. We present a case of a 22-year-old female with multiple papules and nodules in trunk and limbs that after histopathological and immunochemical examination was compatible with Type D LyP. It is important to report this case, as a perfect example of an uncommon variant of LyP, with emphasis in its typical clinical, histopathological and inmunohistochemical findings and review of the literature.

Exploring the IL-21–STAT3 Axis as Therapeutic Target for Sézary Syndrome

Sézary syndrome is an aggressive cutaneous T-cell lymphoma. The malignant cells (Sézary cells) are present in skin, lymph nodes, and blood, and express constitutively activated signal transducer and activator of transcription (STAT)3. STAT3 can be activated by IL-21 in vitro and the IL-21 gene itself is a STAT3 target gene, thereby creating an autocrine positive feedback loop that might serve as a therapeutic target. Sézary cells underwent apoptosis when incubated with Stattic, a selective STAT3 inhibitor. STAT3 activation in Sézary cells did not affect expression of the supposed anti-apoptotic STAT3 target genes BCL2, BCL-xL, and SURVIVIN, whereas expression of (proto)oncogenes miR-21, TWIST1, MYC, and PIM1 was significantly increased. CD3/CD28-mediated activation of Sézary cells induced IL-21 expression, accompanied by STAT3 activation and increased proliferation. Blocking IL-21 in CD3/CD28-activated cells had no effects, whereas Stattic abrogated IL-21 expression and cell proliferation. Thus, specific inhibition of STAT3 is highly efficient in the induction of apoptosis of Sézary cells, likely mediated via the regulation of (proto)oncogenes. In contrast, blocking IL-21 alone seems insufficient to affect STAT3 activation, cell proliferation, or apoptosis. These data provide further insights into the pathogenic role of STAT3 in Sézary syndrome and strengthen the notion that STAT3 represents a promising therapeutic target in this disease.

Abstract 3921: Generation of a T-cell lymphoma mouse model based on chronic LPS challenge and TGF-β signaling disruption

Abstract Background: It has been demonstrated that elevated alcohol intake affects the permeability of the gut allowing endotoxins such as LPS to enter into the portal circulation and to promote liver ethanol toxicity. It has also been shown that increased ethanol intake and LPS challenge impairs liver function and affects liver tumor formation. Hence, with the aim of generating a mouse model of alcoholic liver disease that would rapidly develop steatosis, inflammation and fibrosis, we formulated a regimen that combined chronic injections of low dose LPS with ethanol feeding to mice with impaired TGF-β signaling through constitutive disruption of β2-spectrin (Sptbn1+/-) , Smad3 (Smad3+/-) or both. Methods: Wild type, Sptbn1+/-, Smad3+/- and Sptbn1+/-;Smad3+/- female mice between 8-10 weeks of age were fed orally with a Lieber-DeCarli-based diet containing 6.7% ethanol, and 2-4 weeks later they were injected with 2mg/kg LPS via i.p. 3 times/week for up to 20 weeks or death, whichever occurred first. Survival analysis was done using the Gehan-Breslow-Wilcoxon test. Results: Unexpectedly, the mice treated with chronic low dose LPS and fed the alcohol-containing diet only developed a mild liver phenotype, i.e. steatosis but not hepatitis or fibrosis. Instead, these mice developed very aggressive T-cell lymphomas with massive abdominal and thoracic hemorrhages to which the TGF-β mutant mice succumbed more rapidly than the wild type mice. The median survival of the mutants oscillated between 39 to 42 weeks vs. undefined of the wild type mice. We found statistically significant decreased survival of the mutant mice vs. the wild type (p-values between 0.0015 and 0.028). Conclusions: To our knowledge, this is the first report of a mouse model of aggressive T-cell lymphoma based on chronic challenge with low dose LPS and alcohol feeding. We are currently assessing the cross-talk of the LPS/TLR4 and TGF-β pathways in T-cells from these mice to understand the molecular mechanism underlying the accelerated rate of lymphoma development in these mice. Citation Format: Nina M. Muñoz, Lior H. Katz, Mihai I. Gagea, Asif Rashid, Lopa Mishra. Generation of a T-cell lymphoma mouse model based on chronic LPS challenge and TGF-β signaling disruption. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3921. doi:10.1158/1538-7445.AM2014-3921

Abstract 3719: Mechanisms of resistance to the second-generation alk inhibitor ap26113 in human npm-alk-positive anaplastic large cell lymphoma cells

Abstract ALK is a tyrosine kinase receptor involved in a broad range of solid and haematological tumors. It has been shown that about 70-80% of ALK+ Anaplastic Large Cell Lymphoma (ALCL) cases, an aggressive Non Hodgkin T-cell lymphoma, are characterized by the traslocation t(2;5), originating the functional, aberrant oncogenic fusion protein NPM-ALK. Crizotinib was the first clinically relevant ALK inhibitor, now approved for the treatment of late stage and metastatic cases of lung cancer. Several patients soon developed Crizotinib resistance, mainly due to the appearance of new point mutations located in ALK kinase domain. Currently, other ALK inhibitors are available and already in clinical trial, hopefully representing the second line therapy able to overcome Crizotinib resistance. In this work we focused our attention on the more potent phase I/II dual ALK/EGFR inhibitor AP26113 (Ariad Pharm.), able to overcome the Crizotinib-resistant gatekeeper mutation L1196M. Two NPM-ALK+ human cell lines, KARPAS-299 and SUP-M2, were grown in four independent flasks in the presence of increasing doses of AP26113, so that eight cell lines able to grow in the presence of high AP26113 doses were selected. All cell lines show an AP26113 50% inhibitory concentration (IC50) value substantially higher than the one observed in parental cells, with a 130 to 1000-fold increase. All KARPAS-299 populations resistant to AP26113 show NPM-ALK overexpression, due to oncogene amplification, as the main cause of resistance, while SUP-M2 cells proliferating in the presence of the drug harbour several point mutations spanning the entire ALK kinase domain. In particular, we identified the following aminoacid substitutions: L1122V, , L1196M, S1206C and a double F1174V+L1198F mutation. The knowledge of the possible appearance of new clinically relevant mechanisms of drug resistance is a useful tool for the management of new TKI resistant cases. Citation Format: Monica Ceccon, Luca Mologni, Giovanni Giudici, Rocco Piazza, Alessandra Pirola, Diletta Fontana, Carlo Gambacorti-Passerini. Mechanisms of resistance to the second-generation alk inhibitor ap26113 in human npm-alk-positive anaplastic large cell lymphoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3719. doi:10.1158/1538-7445.AM2014-3719

Allogeneic and autologous stem cell transplantation for hepatosplenic T-cell lymphoma: a retrospective study of the EBMT Lymphoma Working Party.

The objective of this registry study was to analyse the outcome of patients who underwent allogeneic or autologous haematopoietic stem cell transplantation (HSCT) for hepatosplenic T-cell lymphoma (HSTL), a rare and extremely aggressive peripheral T-cell lymphoma subtype. Patients were eligible if they had histologically verified HSTL and underwent HSCT between 2003 and 2011. Seventy-six patients were identified in the European Society for Bone and Marrow Transplantation database. Additional baseline and follow-up information could be obtained from the referring centres for 36 patients. Eleven of these were excluded following histopathology review, leaving 25 patients in the final study cohort (alloHSCT 18, autoHSCT 7). With a median follow-up of 36 months, 2 patients relapsed after alloHSCT, resulting in a 3-year progression-free survival of 48%. After autoHSCT, 5 patients relapsed and subsequently died. This study indicates that graft-versus-lymphoma activity conferred by alloHSCT can result in long-term survival for a substantial proportion of patients with HSTL.

Generation of a mouse model of T-cell lymphoma based on chronic LPS challenge and TGF-β signaling disruption.

asymptomatic steatosis, alcoholic hepatitis and alcoholic cirrhosis, which substantially increase the risk for developing hepatocellular carcinoma. Transforming growth factor (TGF-β) signaling pathway is a major regulator in chronic liver diseases contributing to all liver disease progression from liver injury, inflammation and fibrosis to HCC. With the aim of generating a mouse model of alcoholic liver disease that would rapidly develop steatosis, inflammation as well as fibrosis, we formulated a regimen that combined chronic injections of low dose (2mg/kg) lipopolysaccharide (LPS) with Lieber DeCarli-based diet containing 6.7% ethanol feeding to mice with impaired TGF-β signaling through constitutive disruption of β2-spectrin and/or Smad3. Unexpectedly, the mice treated with chronic low dose LPS and fed the alcohol-containing diet developed very aggressive T-cell lymphomas to which the TGF-β mutant mice succumbed more rapidly than the wild type mice. In contrast, their liver phenotype was mild as they only developed steatosis but not hepatitis or significant fibrosis. To our knowledge, this is the first report of a mouse model of aggressive T- cell lymphoma based on chronic challenge with low dose LPS and TGF-β disruption.

Trial in Progress: Phase 3 Randomized Study of Investigational Drug Alisertib (Mln8237) Vs Investigator'S Choice in Patients (Pts) with Relapsed/Refractory (Rel/Ref) Peripheral T-Cell Lymphoma (Ptcl): the Lumiere Trial

ABSTRACT Background: PTCL is an aggressive form of non-Hodgkin lymphoma (NHL), accounting for 5–10% of NHL diagnoses. Standard NHL therapies developed primarily for B-cell lymphomas are not optimal for PTCL and early relapse is common. Only a minority of pts are eligible for bone marrow transplantation and the majority of transplanted pts will relapse. Approved therapies for rel/ref PTCL in the USA include pralatrexate and romidepsin; elsewhere, gemcitabine is frequently used although not approved for this indication. The oral, investigational drug alisertib is a selective inhibitor of Aurora A kinase (AAK) – a key mitotic regulator overexpressed or amplified in various human tumors. Phase 2 data on single-agent alisertib in rel/ref aggressive T-cell lymphoma (Friedberg et al, J Clin Oncol 2014) support this phase 3 trial in rel/ref PTCL. Trial design: This open-label phase 3 study (NCT01482962) will enroll up to 354 adults with rel/ref PTCL after ≥1 prior systemic cytotoxic therapy at approximately 140 centers in 27 countries worldwide. Pts will be randomized 1:1 to alisertib 50 mg twice daily as an enteric coated tablet on d 1–7 of 21-d cycles, or to investigator's choice of: pralatrexate 30 mg/m2 IV once weekly for 6 weeks in 7-week cycles; romidepsin 14 mg/m2 IV on d 1, 8, 15 of 28-d cycles; or gemcitabine 1000 mg/m2 IV on d 1, 8, 15 of 28-d cycles. Disease subtype must be centrally confirmed. Pts having previously received all comparator drugs are ineligible. Pts will not receive a comparator drug previously received. Crossover to alisertib is not permitted. Primary endpoints are overall response rate and progression-free survival by International Working Group criteria assessed by central review. Secondary endpoints include complete response rate, overall survival, time to progression, time to response, response duration, safety, and quality of life. Exploratory endpoints include candidate biomarker (e.g. AAK, Ki-67) evaluation in tumor biopsies. 205 pts have been randomized as of 15 April 2014. This abstract was accepted and previously presented at the Pan-Pacific Lymphoma Conference in Hawaii on 21/25 July 2014. Disclosure: H. Liu, X. Zhou, C. Dansky Ullmann, V. Kelly and E.J. Leonard: Employment: Takeda; O. O'Connor: Advisory board membership: Takeda Honoraria: Takeda, Seattle Genetics Research funding: Takeda. All other authors have declared no conflicts of interest.

Platelet-derived Growth Factors and Receptors in Canine Lymphoma

Platelet-derived growth factors (PDGFs) belong to a family of polypeptide growth factors that signal through cell surface tyrosine kinase receptors to stimulate growth, proliferation and differentiation. Platelet-derived growth factor receptors (PDGFRs) are also considered important targets for specific kinase inhibitors in the treatment of several human tumours. The aim of this study was to investigate the role of PDGF-A, PDGF-B, PDGFR-α and PDGFR-β in canine lymphoma by determining gene and protein expression in lymph nodes of dogs with diffuse large B-cell lymphoma (DLBCL), peripheral T-cell lymphoma (PTCL), T-lymphoblastic lymphoma (T-LBL) and in healthy control dogs. One lymph node was also studied at the end of therapy in a subset of dogs in remission for DLBCL. In controls, PDGF-A, PDGFR-α and PDGFR-β mRNA levels were significantly higher than in DLBCLs, PTCLs and T-LBLs. However, PDGFR-α and PDGFR-β were minimally expressed by lymphocytes and plasma cells in normal lymph nodes as determined by immunohistochemistry, while neoplastic B and T cells showed the highest score (P <0.05). This discordant result may be compatible with the constitutive expression of these molecules by endothelial cells and fibroblasts in normal lymph nodes, thereby influencing gene expression results. Furthermore, these cells were not included in the immunohistochemical analysis. Similarly, dogs with DLBCL that were in remission at the end of therapy showed significantly higher gene expression of PDGFs and receptors than at the time of diagnosis and with an opposite trend to the protein assay. PDGF-B protein and mRNA were overexpressed in PTCLs and T-LBLs when compared with DLBCLs and controls (P <0.05). Additionally, there was a correlation between protein expression of PDGF-B and both PDGFRs in PTCLs and T-LBLs, suggesting an autocrine or paracrine loop in the aetiology of aggressive canine T-cell lymphomas. These data provide a rationale for the use of PDGFR antagonists in the therapy of aggressive T-cell lymphomas, but not in DLBCLs.