- Maria Pamela Dobay1,
- Francois Lemonnier2,3,
- Edoardo Missiaglia1,4,
- Christian Bastard5,
- David Vallois4,
- Jean-Philippe Jais6,
- Laurianne Scourzic7,
- Aurélie Dupuy2,3,
- Virginie Fataccioli2,3,14,
- Anais Pujals2,3,14,
- Marie Parrens8,
- Fabien Le Bras9,
- Thérèse Rousset10,
- Jean-Michel Picquenot11,
- Nadine Martin2,3,
- Corinne Haioun2,3,9,
- Richard Delarue12,
- Olivier A. Bernard7,
- Mauro Delorenzi1,13,
- Laurence de Leval4,*⇑ and
- Philippe Gaulard2,3,14,*⇑
- 1SIB Swiss Institute of Bioinformatics, Lausanne, Switzerland
- 2INSERM U955, Créteil, France
- 3Université Paris-Est, Créteil, France
- 4Institut de Pathologie, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
- 5Laboratoire de cytogénétique et biologie moléculaire, CLCC H. Becquerel, Rouen, France
- 6Service de Biostatistiques, GH Necker Enfants Malades, Paris, France
- 7Institute Gustave Roussy, Villejuif, France
- 8Departement de Pathologie, CHU de Bordeaux, Hopital du Haut Lévêque, Pessac, France
- 9Unité Hémopathies lymphoïdes, Hôpital Henri Mondor, AP-HP, Créteil, France
- 10Service d’anatomo-pathologie, CHU Gui de Chauliac, Montpellier, France
- 11Service d’anatomo-pathologie, CLCC H. Becquerel, Rouen, France
- 12Hématologie Clinique, GH Necker-Enfants malades, Paris, France
- 13Ludwig Center for Cancer Research, University of Lausanne, Switzerland
- 14Hôpital Henri Mondor, Département de Pathologie, AP-HP, Créteil, France
- Correspondence: /
In addition to angioimmunoblastic T-cell lymphoma (AITL), the 2016 revised WHO classification of haematological malignancies recognizes two provisional lymphoma entities of follicular helper T-cell (TFH) derivation, namely follicular peripheral T-cell lymphoma (F-PTCL) and nodal PTCL with a TFH phenotype.1 Here, we performed a comprehensive, integrative clinicopathological and molecular analysis of these three entities. We found that F-PTCL and other nodal PTCL with TFH phenotype share not only immunophenotypical features, but also similar clinical, genetic and molecular features with AITL. Our results support the view that these lymphomas belong to the spectrum of a common disease.
AITL and PTCL, not otherwise specified (PTCL-NOS), account for the majority of nodal PTCLs.2,3 While PTCL-NOS is by definition heterogeneous and an exclusion diagnosis, AITL is characterized by a constellation of clinical, morphological, and immunophenotypical features, and defined by its cellular derivation from TFH cells.1,4 The typical pathological features, i.e., clear cells, increased vascularization, follicular dendritic cell (FDC) proliferation, and the presence of eosinophils, inflammatory cells, and EBV-positive B-blasts, are variably developed.5,6
An overlap between AITL and PTCL-NOS was substantiated by the observation that a subset of cases diagnosed as PTCL-NOS, upon routine pathological evaluation, actually harbor imprints of the TFH signature4 and/or express TFH-associated markers.7,8 Furthermore, the rare “follicular” T-cell lymphoma (F-PTCL), initially classified as a PTCL-NOS variant, is characterized by a TFH immunophenotype and clinicopathological features overlapping with AITL.9,10 Recently, recurrent AITL-associated TET2, DNMT3A and RHOA mutations were also found in a subset of PTCL-NOS, and tended to correlate with TFH features.11,12 Based on these recent findings, the 2016 update of the WHO classification groups AITL and other nodal lymphomas of TFH origin under the same umbrella.1 However, a thorough, systematic and multiparametric comparison of these entities is lacking.
Here, we performed a comprehensive integrative clinicopathological and molecular analysis comparing AITL and other nodal PTCLs of TFH origin. Twenty-one such cases, including five F-PTCL, were identified in the TENOMIC biobank of the LYSA, and compared to 94 AITL and 36 PTCL-NOS. All cases were reviewed by three hematopathologists according to the 2016 WHO classification criteria (1 and Online Supplementary Information). The five F-PTCL by definition comprised FDCs associated to the follicles without extrafollicular FDC expansion, and were positive for all TFH markers tested (4 or 5) (Figure 1A–B). The 16 other nodal PTCL with TFH phenotype (referred to as “TFH-like PTCL”) (Figure 1A, 1C, 1D) lacked typical morphological AITL features, seven of them disclosed some FDC expansion, and all expressed a TFH phenotype defined by expression of at least two TFH markers among ICOS (14/15, 93%), PD1 (8/10, 80%), CXCL13 (11/15, 73%), BCL6 (9/13, 69%) and CD10 (6/16, 37.5%). Overall, TFH-like PTCL cases tended towards a more incomplete TFH profile compared to AITL (Figure 1A, Online Supplementary Table S1); PTCL-NOS cases included cytotoxic PTCLs, non-cytotoxic PTCLs strongly positive for CD30, and others with a non-TFH immunophenotype (Online Supplementary Figure S1). All five F-PTCL and 13/16 TFH-like PTCL comprised scattered CD20+ and/or EBV-positive blasts (Figure 1A).
Clinically, TFH-like and F-PTCL patients had a high incidence of disseminated disease (93% and 100%, respectively), closer to the incidence in AITL (99%) than PTCL-NOS cases (85%, P<0.02). Within the limits of available data, other AITL-related features, such as a positive Coombs test and anemia, also occurred more frequently in TFH-PTCL than in PTCL-NOS (Table 1). Overall survival compared across entities using the Kaplan-Meier and log-rank methods did not significantly differ (Online Supplementary Figure S2), consistent with previous reports of poor outcomes (30–35% 5-year OS) for AITL and PTCL-NOS.2,3
Targeted sequencing performed in 114 cases, including 85 previously reported,11,13 revealed similar mutation frequencies in TET2, DNMT3A and RHOA in AITL and other nodal lymphomas of TFH origin, while IDH2 mutations, with a single exception, were restricted to AITL, consistent with recent findings (Figure 1A, Table 1). The RHOA G17V mutation rate in TFH-like (57%) and F-PTCL (60%) was comparable to the 58% frequency found in AITL.12 TET2 mutations were even more frequent in TFH-like (64%) and F-PTCL (75%) than in AITL (48%). The mutational portrait of F-PTCL has not yet been reported. Here, we found that 3/5 F-PTCLs carried RHOA and TET2 mutations, plus a DNMT3A mutation in one case (Figure 1A). ITK-SYK translocations occurred in 2/5 F-PTCLs, one of which was TET2-mutated, and the other wild-type for the four genes tested. The frequency of mutations in PTCL-NOS was significantly lower (TET2, 17%; RHOA G17V, 0%; DNMT3A, 4%; IDH2, 0%). These results extend previous findings, demonstrating overlapping genetic characteristics in AITL and other PTCL of TFH origin,13 including F-PTCL.
To investigate the clustering of F-PTCL and TFH-like PTCL compared to AITL and PTCL-NOS, we performed principal component analysis (PCA) of the top 6000 most variably expressed genes based on gene expression profiles of 144 cases.4 We calculated pairwise distances between cluster centroids (ΔCcentroid) and cluster dispersion (D), measured by the distance of each sample from its cluster centroid, in the PCA space. Differences in D were evaluated using a one-tailed t-test. By PCA of GEP (Figure 1E, Online Supplementary Figure S3A), AITL was more homogenous (D=0.07) than PTCL-NOS (D=0.13). Other lymphomas of TFH origin clustered closer to AITLs (ΔCcentroid=0.04) than to PTCL-NOS (ΔCcentroid=0.10). AITL and other lymphomas of TFH origin (D=0.1) were also significantly less dispersed (P-value=0.03) than PTCL-NOS across the 3D principal component space. Interestingly, F-PTCLs were completely contained within AITLs, and were similarly dispersed (Figure 1E, Online Supplementary Figure S3A).
Focusing on molecular signatures characteristic of AITL,4 the median level of TFH signature expression in PTCLs of TFH origin (7.33) was comparable to that in AITL (7.41) (P=0.47) (Figure 1F), further supporting that the expression of at least two TFH-associated molecules captures a TFH signature. In contrast, the AITL signature and AITL microenvironment signatures were both expressed at lower levels in PTCL of TFH origin than in AITL on average, but higher than in PTCL-NOS (P<0.01 for all pairwise comparisons), consistent with the lack of the characteristic AITL microenvironment in other PTCLs of TFH origin. Notably, the expression range of the AITL signature in PTCL of TFH origin was distributed within the observed range for AITL, except in one sample. We also performed pairwise gene set enrichment analyses using key biological signatures. Our results reinforce similarities between AITL and PTCL of TFH origin in terms of the expression of TFH lineage pathways, key signaling pathways such as JAK-STAT and IL2/IL17/IL15-mediated cytokine signaling as well as several transcription targets, including NFKB- and STAT3-induced genes (Online Supplementary Figure S3B–C, Online Supplementary Table S3). Altogether, the striking overlap of AITL, F-PTCL and TFH-like PTCLs in both global and specific gene expression patterns, suggests that these lymphomas, despite morphological differences, belong to the same spectrum. Interestingly, it was proposed that TFH-like PTCL with expanded FDC meshwork could represent a “tumor cell-rich” variant of AITL, as in seven of our patients.5
After correction for tumor content differences (Online Supplementary information), array CGH data showed a similar frequency of samples with events in AITL (23/60, 38%) and other PTCL of TFH origin (5/15, 33%), whereas a much higher proportion of PTCL-NOS (17/27, 63%) had abnormal profiles. There was, however, an increasing frequency of cases with heavy rearrangements and/or homozygous deletions or amplifications from AITL to other TFH-PTCL to PTCL-NOS (Table 1). In addition, AITL-linked copy number gains14 in chromosomes 5 (9/60 AITL, 15%) and 7 (5/60 AITL, 8 %) occurred with similar frequency in other PTCL of TFH origin (1/15, 7%; and 2/15, 13%, respectively) (Online Supplementary Figure 4). Gene losses likely linked to secondary oncogenic events that potentially correlate with gene expression are given in Online Supplementary Table S4.
In conclusion, our results show that F-PTCL and nodal PTCL with TFH phenotype, as defined in the updated WHO classification,1 share with AITL not only phenotypic features, but also similar clinical features, genetic events and molecular signatures. Our results therefore support the dissociation of F-PTCL and PTCL with TFH-phenotype out of the PTCL-NOS entity, and the grouping of these new provisional entities together with AITL, based on their TFH cell derivation as the unifying feature.1 It is recommended that these lymphomas of TFH origin be identified by routinely investigating any PTCL-NOS for TFH-associated marker expression and AITL-like features and, when possible, key AITL-associated genetic lesions. While the distinction between PTCL of TFH origin versus PTCL-NOS has currently no impact on clinical management, the situation could change in the near future with the introduction of new therapeutic approaches and development of targeted interventions, more or less likely to be operant in one or the other category, such as hypomethylating agents.15 Our results also support that AITL, F-PTCL and PTCL with TFH phenotype truly represent variations along the spectrum of a single disease entity. Whether the distinction between the prototypic TFH neoplasm (AITL) and the two less prevalent provisional entities needs to be maintained in the future remains an open question which should be addressed by prospective observational studies.
The authors thank Yves Allory from the Tumor Tissue Bank (Plateforme de Ressources Biologiques, GH H. Mondor, AP-HP, Créteil, France) and the LYSA-Pathology from the LYSARC for its technical and logistical assistance.
↵* equal contributions
Funding: this work was supported by grants from the MEDIC Foundation, the Ligue du Cancer (Switzerland), Plan Cancer (Belgium), the Institut National du Cancer (INCa AAP PLBIO13-085and INCa-Plan Cancer 2013), the Fondation pour la Recherche Médicale, the Cancéropole Ile-de-France, and ARTGIL.
Information on authorship, contributions, and financial & other disclosures was provided by the authors and is available with the online version of this article at www.haematologica.org.
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