Differential SATB1 Expression Reveals Heterogeneity of Cutaneous T-Cell Lymphoma

SATB1 is an important T-cell specific chromatin organizer in cutaneous lymphoma, whereas its expression and function mycosis fungoides (MF) remain ambiguous. Our study aimed to investigate the clinicopathological significance of a cohort 170 patients with MF. was heterogeneous among MF each clinical stage. High associated epidermal hyperplasia, eosinophil infiltration, less large-cell transformation, favorable prognosis cases. CD30 coexpression distinguished CD30+ lymphoproliferative disorders from transformation. silencing lines showed that upregulated genes involved recruitment, including signal transducer activator transcription 3 IL13, downregulated cell-cycle progression, which may explain inferior for low SATB1-expressing Moreover, inversely correlated PD-1 expression, indicating exhausted status SATB1-negative malignant T cells. positively toll-like receptors suggesting innate immune activation high Therefore, variable promotes heterogeneity pathology outcome Mycosis most common lymphoma (CTCL) comprises about 50% primary lymphomas (Willemze et al., 2005Willemze R. Jaffe E.S. Burg G. Cerroni L. Berti E. Swerdlow S.H. al.WHO-EORTC classification lymphomas.Blood. 2005; 105: 3768-3785Crossref PubMed Scopus (3090) Google Scholar). Whereas at early stage have indolent course prognosis, up 20% experience disease progression and/or death within 10 years (Agar 2010Agar N.S. Wedgeworth Crichton S. Mitchell T.J. Cox M. Ferreira al.Survival outcomes prognostic factors fungoides/Sézary syndrome: validation revised International Society Cutaneous Lymphomas/European Organisation Research Treatment Cancer staging proposal.J Clin Oncol. 2010; 28: 4730-4739Crossref (509) The mechanisms underlying development largely undetermined. thymocyte-specific plays critical role (Alvarez 2000Alvarez J.D. Yasui D.H. Niida H. Joh T. Loh D.Y. Kohwi-Shigematsu MAR-binding protein orchestrates temporal spatial multiple during development.Genes Dev. 2000; 14: 521-535PubMed Scholar) differentiation (Burute 2012Burute Gottimukkala K. Galande Chromatin determinant differentiation.Immunol Cell Biol. 2012; 90: 852-859Crossref (25) Recent studies our group others demonstrated aberrations CTCLs. Loss found aggressive form CTCL (Sézary syndrome) (Wang 2011Wang Y. Su Zhou L.L. Tu P. Zhang X. Jiang al.Deficiency Sezary cells causes apoptosis resistance by regulating FasL/CD95L transcription.Blood. 2011; 117: 3826-3835Crossref (46) Scholar), shown portion CTCLs, (Fredholm 2018Fredholm Willerslev-Olsen A. Met Ö. Kubat Gluud Mathiasen S.L. al.SATB1 cells.J Invest Dermatol. 2018; 138: 1805-1815Abstract Full Text PDF (29) Scholar; Grzanka 2015Grzanka D. Gagat Izdebska Marszałek Expression special AT-rich sequence-binding 1 independent factor lymphoma.Oncol Rep. 2015; 33: 250-266Crossref (12) Scholar, 2012Grzanka Tadrowski Sokołowska-Wojdyło al.Correlation lymphomas.Pol J Pathol. 63: 101-105PubMed CD30-positive (CD30+LPDs) (Sun 2018Sun J. Yi Qiu Fu W. Wang Liu F. defines subtype T-helper 17 cytokine profile.J 1795-1804Abstract (19) breast implant‒associated anaplastic (BIA-ALCL) (Kadin 2016Kadin M.E. Deva Xu Morgan Khare MacLeod R.A.F. al.Biomarkers provide clues events pathogenesis implant-associated large cell lymphoma.Aesthet Surg 2016; 36: 773-781Crossref (100) In CD30+LPDs, diseases helper type profile better response treatment MF, discrepant multifaceted functions (Gagat 2019Gagat Krajewski Ambiguous tumors.J 2019; 139: 1608-1610Abstract (4) Poglio Merlio, 2018Poglio Merlio J.P. pivotal epigenetic biomarker lymphomas.J 1694-1696Abstract (9) Stage-related decrease relation unfavorable been small cohorts However, correlation between features utility settings are not fully determined. mechanism discrepancy stage-dependent remains unknown. this study, we characterized identified intrinsic corresponding To comprehensively evaluate performed immunostaining on 155 lesional biopsies 145 those patch (n = 30), plaque 58), tumor 57). infiltrating lymphocytes varied totally negative staining nearly completely positive (Figure 1a). Because it difficult distinguish reactive histology, especially stages, levels were evaluated according percentages all lymphocytes. A total designated into two groups basis expression: SATB1-high (SATB1-H, n 100) least 40% SATB1-low (SATB1-L, 45) <40% SATB1-positive Stage-specific analysis 77.3% patch- plaque-stage cases only 57.9% samples had 1b), stage-related as reported previously This result further validated ImageJ IHC Profiler (Varghese 2014Varghese Bukhari A.B. Malhotra De Profiler: open source plugin quantitative evaluation automated scoring immunohistochemistry images human tissue samples.PLoS One. 2014; 9: e96801Crossref (386) quantification system evaluating nuclear staining. Quantification results four-point decreased compared 1c). 22.7% stages or loss SATB1, wonder whether resulted gradual tendency toward progression. expressions who follow-up examined. consistent 1d). Among them, 4 5 experiencing SATB1-L These suggested existed different propensity characterize retrospectively reviewed information these patients. duration comparable SATB1-H (Table 1). gender, age onset 1), variants 2a). predictive pruritus, modified severity-weighted assessment tool scores, before developing overt tumors (Olsen 2011Olsen E.A. Whittaker Kim Y.H. Duvic Prince H.M. Lessin S.R. al.Clinical end points criteria Sézary consensus statement Lymphomas, United States Lymphoma Consortium, Task Force European Cancer.J 29: 2598-2607Crossref (397) Kaplan‒Meier survival progression-free worse 2b). exclude effect early-stage (IA–IIA) advanced-stage (IIB–IVB) analyzed separately. Progression-free analyses both advanced revealed respect mRNA 49 confirmed related (Supplementary Figure S1).Table 1Clinical Pathological Features Patients Varied ExpressionVariableSATB1-L, (%)SATB1-H, (%)Total, nP-valueGender1450.218 Male24 (54.5)66 (65.3) Female20 (45.5)35 (34.7)Age onset1450.832 Mean ± SD39.6 17.839.2 16.0Follow-up (mo)1450.595 (95% CI)23.7 (18.6–28.7)22.9 (19.4–26.3)Clinical characteristics Pruritus1450.338Absent15 (34.1)43 (42.6)Present29 (65.9)58 (57.4) mSWAT571Only naive included.0.767Mean CI)43.0 (21.7–64.3)36.5 (27.1–45.8) Duration (mo)2According patient history.600.134Mean CI)69.9 (47.6–92.2)92.4 (70.2–114.7)Pathological Epidermal hyperplasia1433Epidermis present slides 143 cases.0.0374Statistically significant.Absent24 (55.8)37 (37.0)Present20 (44.2)62 (63.0) Epidermotropism1433Epidermis cases.0.106Absent0 (0.0)8 (8.0)Present44 (100.0)92 (91.2) Ulceration1450.492Absent40 (90.9)95 (94.1)Present4 (9.1)6 (5.9) LCT1450.0004Statistically significant.No22 (50)80 (79.2)Yes22 (50)21 (20.8) Eosinophil infiltration1450.0494Statistically significant.Grades 0–137 (84.1)69 (68.3)Grades 2–37 (15.9)32 (31.7) Ki-67, %925Only Ki-67 data available included.0.074Mean CI)39.5 (27.7–51.2)27.9 (21.5–34.3)Abbreviations: CI, confidence interval; LCT, transformation; mSWAT, tool; SATB1-H, SATB1-high; SATB1-L, SATB1-low.1 Only included.2 According history.3 Epidermis cases.4 Statistically significant.5 included. Open table new tab Abbreviations: SATB1-low. hyperplasia more frequently observed Large-cell transformation (LCT) significantly frequent (50.0% vs. 20.8%, P 0.000), supporting association aggressiveness. Consistently, index higher marginal (39.5% 27.9%, 0.074). Of note, infiltration lesions 2c), classified 0–3 grades 2018Kadin Epstein A.L. Sieber Hubbard B.A. al.IL-13 produced lymphoma: implications pathogenesis.Hum 78: 54-62Crossref (38) Significant correlations well tumor-stage‒only 2d). characteristic histology features, increased LCT. LCT highly representative significant CD30+LPDs raised question can help differentiate CD30-rich MF-LCT currently. developed nodules lesions. tumorous nodular evaluated. patients, five them absent staining, remaining 12 3a). immunofluorescence double colocalization seven other separate costaining cells, spontaneous regression occurred diagnosis secondary CD30, experienced required treatment, 3b). suggest favors over MF-LCT. oncogenic somatic mutations JAK1 (STAT)3 5% (Crescenzo 2015Crescenzo Abate Lasorsa Tabbo’ Gaudiano Chiesa N. al.Convergent kinase fusions lead STAT3 lymphoma.Cancer Cell. 27 ([published correction appears 2015;27:744]): 516-532Abstract (268) prompted us possible mutation expression. Tissue DNA 15 14 (five four MF-LCT, lymphomatoid papulosis, [PCALCL], one papulosis PCALCL) above sequenced hotspot PCALCL Maurus 2020Maurus Appenzeller Roth Brändlein Kneitz Goebeler al.Recurrent JAK STAT alterations [e-pub ahead print].J 2020; (accessed May 2020)https://doi.org/10.1016/j.jid.2020.02.019Abstract Ohgami 2013Ohgami R.S. Ma Merker Martinez B. Zehnder J.L. Arber D.A. granular lymphocytic leukemia.Leukemia. 2013; 27: 2244-2247Crossref (56) No nonsynonymous detected, mutation. Hence, serve valuable marker CTCL, illustrated algorithm proposed 3c). explore mechanism(s) silenced MJ highest Transcriptome sequencing following Kyoto Encyclopedia Genes Genomes pathway glycosaminoglycan biosynthesis, cytokine‒cytokine receptor interaction, JAK-STAT signaling cycle, replication, transcriptional misregulation cancer 4a Supplementary Table S1). encoding 2 IL13 4b d). confirm result, Myla transduced lentiviral vector SATB1. IL-13 overexpression 4c e), upregulated, be due constitutional (McKenzie 2012McKenzie R.C. Jones C.L. Tosi I. Caesar J.A. S.J. Constitutive syndrome SHP-1.Leukemia. 26: 323-331Crossref (23) validate regulatory relationship lesions, tumor-stage expressed much 4f).Figure 4SATB1 regulates signaling, cycle. (a) KEGG differentially (ordered enrichment score, blue font: pathways enriched genes; red genes). (b) suppression shRNA sequences (sh1 sh2) targeting Cells scrambled (sh0) controls. ∗P < 0.05, ∗∗∗P 0.001. (c) SATB1-overexpressing Parental empty (LV-0) (d) level sh0 (e) LV-0 (f) differential levels. 0.05. 49, unpaired Student’s t-test. (g) Spearman grade tumors. (h) Pearson CCL11, CCL24, CCL26, (i) Upregulated (j) E2F1 MCM3 ∗∗P 0.01, Con, control; FPKM, fragments per kilobase transcript million mapped reads; KEGG, Genomes; fungoides; ns, significant; SATB1-low; sh, short hairpin; STAT, transcription.View Large Image ViewerDownload Hi-res image Download (PPT) recruiting eosinophils (Matucci 2019Matucci Maggi Vultaggio Eosinophils, IL-5/IL-5Rα axis, biologic effects benralizumab severe asthma.Respir Med. 160: 105819Abstract (21) Neilsen Bryce, 2010Neilsen C.V. Bryce P.J. Interleukin-13 directly oesophagus production CCL11 CCL24 migration eosinophils.Clin Exp Allergy. 40: 427-434Crossref Soroosh Doherty, 2009Soroosh Doherty T.A. Th9 allergic disease.Immunology. 2009; 127: 450-458Crossref (185) accordance that, 4g), responsible recruitment recruits through eotaxin-1 (CCL11), eotaxin-2 (CCL24), eotaxin-3 (CCL26), potent attractants (Larose 2015Larose M.C. Chakir Archambault A.S. Joubert Provost V. Laviolette CCL26 bronchial epithelial airway eosinophils: involvement eosinophilic asthma.J Allergy Immunol. 136: 904-913Abstract (49) As expected, 4h). also S2). Meanwhile, series regarding cycle E2F1, CCNB2, ORC1, CDC45, MCM3, MKI67 4i). An has 4j). regulated contributed negatively line Growing evidence indicates upstream suppressor (Stephen 2017Stephen T.L. Payne K.K. Chaurio R.A. Allegrezza M.J. Zhu Perez-Sanz governs repression tumor-reactive cells.Immunity. 2017; 46: 51-64Abstract (75) Yasuda 2019Yasuda Kitagawa Kawakami Isaka Watanabe Kondoh al.Satb1 effector program encephalitogenic Th17 chronic inflammation.Nat Commun. 10: 549Crossref (11) hallmark cases, (Cetinözman 2012Cetinözman Jansen P.M. Vermeer M.H. Willemze Differential programmed death-1 (PD-1) fungoides.Arch 148: 1379-1385Crossref (76) 2015Wang Ni Covington K.R. Yang B.Y. Shiu al.Genomic profiling identifies key genes.Nat Genet. 47: 1426-1434Crossref (202) Given remarkable intriguing see suppress none express PD-1, neither quiescent state nor under stimulation phytohemagglutinin, superantigen well-known upregulate S3). Thus, immunostainings reveal Membrane quantified software module Visiopharm (Brügmann 2012Brügmann Eld Lelkaitis Nielsen Grunkin Hansen al.Digital membrane connectivity robust measure HER2 immunostains.Breast Res Treat. 132: 41-49Crossref defined score number intensity than 5a b). minority 5c) PD-1–positive larger lymphocytes, Furthermore, based p