Team 1: Physiopathology of cutaneous T cell lymphomas and receptors of innate immunity (Team leader: Armand Bensussan)

 Projects: (recent publications below)


1. KIR3DL2/CD158k, a potential therapeutic target through the use of monoclonal antibody

A therapeutic strategies being developed in a number of hematological malignancies is to mobilize the innate immunity of patients by injection of monoclonal antibodies. Thus, alemtuzumab (anti-CD52 antibody) was used on patients with Sézary syndrome, leading to a rate of full or partial remission of 38%. However, hematological and immunosuppressive toxicity of the antibody was observed, mainly due to the fact that the induced cytotoxic activity does not distinguish between normal lymphocytes and malignant cells, causing profound leukopenia and infectious complications. Numerous studies in the laboratory have demonstrated the validity of KIR3DL2/CD158k as diagnostic and prognostic marker in Sézary Syndrome.

A research project is being conducted in collaboration with the company Innate Pharma (Marseille, France), to assess the efficacy of humanized chimeric anti-KIR3DL2/CD158k antibodies to induce a NK cell-dependent cytotoxicity. The realization of this work should determine whether KIR3DL2/CD158k could be used as target in order to promote an effective and specific elimination of tumor cells.


2. KIR3DL2/CD158k, a novel receptor for ODN CpG: functionnal and therapeutic implications

Recently, CpG ODN have identified as a new ligand has been identified for KIR3DL2/CD158k. In a collaborative, we demonstrated the specific binding of CpG ODN on KIR3DL2+ NK cells, this interaction leading to partial or complete internalization of the receptor according to the type of CpG ODN used (A, B or C, ODN-C inducing a complete disappearance of KIR3DL2 from the cell surface). Co-localization with TLR9, CpG ODN receptor presenting exclusive endosomal localization, is then observable (Sivori S. et al., Blood 2010). From a functional standpoint, the CpG ODN induce the secretion of IFNg and TNFα by KIR3DL2+ NK cells, suggesting an important role for this receptor in the NK cell responses mediated by CpG ODN. The identification of KIR3DL2 as a receptor for CpG ODN raises the question of their potential effects on the mechanisms controlling proliferation and/or resistance to apoptosis of Sézary cells. We have already established that incubation of Sézary cell lines, or circulating tumor cells from peripheral blood of Sezary patients, with a C-type CpG ODN led to a partial or total disappearance of KIR3DL2 from the cell surface.  We are now working on the functional consequences of such KIR3DL2 engagement in Sézary cells. The results that we will obtain for this part of the project could provide new answers on the pathophysiology of Sézary syndrome, including the factors triggering a tumor proliferation. In addition, validation of a model where KIR3DL2 would ensure the transport of CpG ODNs to their receptor at the intracellular level could afford to consider an immunotherapy based on anti-KIR3DL2 antibody able to block its interaction with CpG ODN.

 

3. Study of new phenotypic markers of Sézary cell, TWIST

Besides the T-plastin, we focus on another factor identified by transcriptome studies of CTCL, the transcription factor TWIST, for two main reasons: its gene expression is regulated by NF-kB in mammalian cells and conversely, it can also regulate the NF-kB pathway, which may contribute to its potential oncogenic role. This embryonic TWIST gene is frequently reactivated in human cancers such as carcinomas, melanoma and osteosarcoma, and it would promote the growth of solid tumors by blocking p53 and inhibiting the apoptosis induced by c-myc. Molecular data suggest that induction of expression of TWIST by NF-kB plays an important role in the resistance to cytotoxic drugs due to the NF-kB. This leads us to study the link between the constitutive activation of NF-kB and TWIST overexpression in tumor cells of CTCL, given their chemoresistance associated with NF-kB, as we have previously demonstrated. A recent study also reported an increased expression of TWIST in lesions of advanced stages of MF/SS, suggesting a correlation between TWIST overexpression and disease progression. Our first results can assess that the synthesis of TWIST in CTCL is controlled by NF-kB  pathway and that this factor is involved in the resistance to apoptosis of tumor cells.

 

4. Identification of CD160TM ligands and of the signalling pathways generated by its engagement at the surface of normal NK lymphocytes and NK/T lymphomas

We identified a novel transmembrane isoform of the receptor CD160 (CD160-TM) only on activated NK cells, NK leukemia and NK/T lymphoma. The total identity of the extracellular domain of CD160 and CD160-TM suggests that the two molecules could interact with the same ligand. However, we observed that the monoclonal antibody BY55 recognizes only CD160, and that anti-CD160-TM antibodies that we produced are unable to interact with the GPI form of the receptor. It is therefore likely that the replacement of the GPI anchor by a transmembrane domain and an intracellular portion results in conformational changes of the extracellular domain, resulting in differential recognition of two molecules by the same antibody. Such a phenomenon has been observed for example for CD16. Although we observed an association of CD160-TM with the HLA-C molecules (ligand for CD160), interaction with other potentental ligands will be investigated. The structural differences between CD160 and its transmembrane isoform raises the question of the intracellular signalling established after recruitment of each receptor. In addition the presence of an intracellular domain in CD160-TM offers the possibility of a signalling pathway distinct from that corresponding to CD160. Our initial results suggest that once recruited, the intracellular domain of CD160-TM is tyrosine phosphorylated by p56lck, this modification being required for the activation of Erk. The phosphorylation motif present in the intracellular domain of CD160-TM, and identified as essential for the establishment of an activating signal (Y225), does not match any consensus sequence described as allowing an association with the SH2 domain of molecules of classical signalling. Consequently, the identity of the molecules relaying the signals generated after engagement of CD160-TM will be sought. Comparison of protein profiles associated with the wild-type protein or a Y225 mutant should allow the identification of intracellular partners necessary for the delivery of an activation signal from CD160-TM. We also seek to establish the functional consequences of CD160-TM engagement on the surface of NK cells in terms of cytokine production, to determine whether the recruitment of CD160-TM led to the mobilization of activation pathways distinct from those involved during CD160 engagement. Finally, the recent characterization of a monoclonal antibody specific for CD160-TM should allow us to identify the intracellular signals attributable to CD160 and CD160-TM when expressed simultaneously on the surface of NK cells.

 

5. Expression of CD160-GPI by normal cutaneous CD4+ T lymphocytes

We do not confirm the results published by others indicating that CD160 is present on the surface of all CD4+ peripheral blood T lymphocytes. We show with the reference anti-CD160 monoclonal antibody, BY55, that only 0.5 to 1% of circulating CD4+ T lymphocytes expresses CD160. This percentage increases during cell activation ex vivo. Interestingly we have shown that CD4+ CD160+ T cells are present in healthy and inflammatory skin. Recently, we were interested to lymphocytes of normal skin and skin biopsies of CTCL. In blood, the CD4+ CD160+ are predominantly CD8aa+ and are cytotoxic. In healthy skin and that of mycosis fungoides and Sézary syndrome, these cells represent between 30 to 60% of T lymphocytes. Our project is to functionally study these cutaneous T cells and to determine the molecular characteristics that distinguished them from peripheral blood CD4+ T lymphocytes.

Recent publications (2014-2016)

 

2016

1. Chasset F, Le Buanec H, Sicre de Fontbrune F, de Masson A, Rivet J, Bergeron A, Bagot M, Socié G, Bensussan A, Bouaziz JD. Evidence of Th1, Th17 and Tc17 cells in psoriasiform chronic graft-versus-host disease. Exp Dermatol, 2016 Jan;25(1):64-5. doi: 10.1111/exd.12857. Epub 2015 Nov 2. No abstract available.

 

2. Brun C, Jean-Louis F, Oddos T, Bagot M, Bensussan A, Michel L. Phenotypic and functional changes in dermal primary fibroblasts isolated from intrinsically aged human skin. Exp Dermatol, 2016 Feb;25(2):113-9. doi: 10.1111/exd.12874. Epub 2016 Jan 11.

 

3. Hu HH, Kannengiesser C, Lesage S, André J, Mourah S, Michel L, Descamps V, Basset-Seguin N, Bagot M, Bensussan A, Lebbé C, Deschamps L, Saiag P, Leccia MT, Bressac-de-Paillerets B, Tsalamlal A, Kumar R, Klebe S, Grandchamp B, Andrieu-Abadie N, Thomas L, Brice A, Dumaz N, Soufir N. PARKIN Inactivation Links Parkinson's Disease to Melanoma. J Natl Cancer Inst, 2015 Dec 17;108(3). pii: djv340. doi: 10.1093/jnci/djv340. Print 2016 Mar.

 

4. Hurabielle C, Michel L, Ram-Wolff C, Battistella M, Jean-Louis F, Beylot-Barry M, d'Incan M, Bensussan A, Bagot M. Expression of Sézary Biomarkers in the Blood of Patients with Erythrodermic Mycosis Fungoides. J Invest Dermatol, 2016 Jan;136(1):317-20. doi: 10.1038/JID.2015.360.

 

5. Lédée N, Petitbarat M, Chevrier L, Vitoux D, Vezmar K, Rahmati M, Dubanchet S, Gahéry H, Bensussan A, Chaouat G. The Uterine Immune Profile May Help Women With Repeated Unexplained Embryo Implantation Failure After In Vitro Fertilization. Am J Reprod Immunol, 2016 Jan 18. doi: 10.1111/aji.12483. [Epub ahead of print

 

6. Joffre J, Hau E, Zeboudj L, Laurans L, Battistella M, Boufenzer A, de Masson A, Le Buanec H, Cochaud S, Bagot M, Bensussan A, Gibot S, Bouaziz JD, Ait-Oufella H. Trem-1 is not crucial in psoriasiform imiquimod-induced skin inflammation in mice. Exp Dermatol, 2016 Jan 19. doi: 10.1111/exd.12944. [Epub ahead of print]

 

7. Boonk SE, Zoutman WH, Marie-Cardine A, van der Fits L, Out-Luiting JJ, Mitchell TJ, Tosi I, Morris SL, Moriarty B, Booken N, Felcht M, Quaglino P, Ponti R, Barberio E, Ram-Wolff C, Jäntti K, Ranki A, Bernengo MG, Klemke CD, Bensussan A, Michel L, Whittaker S, Bagot M, Tensen CP, Willemze R, Vermeer MH. Evaluation of immunophenotypic and molecular biomarkers for Sézary syndrome using standard operating procedures: multicenter study of 59 cases. J Invest Dermatol, 2016 Feb 27. doi: 10.1016/j.jid.2016.01.038.

 

8. Laugier F, Delyon J, André J, Bensussan A, Dumaz N. Hypoxia and MITF regulate KIT oncogenic properties in melanocytes. Oncogene. 2016 Mar 14. doi: 10.1038/onc.2016.39. [Epub ahead of print]

 

9. Chasset F, De Masson A, Le Buanec H, Xhaard A, Sicre de Fontbrune F, Robin M, Rybojad M, Parquet N, Brignier AC, Coman T, Bengoufa D, Bergeron A, Peffault de Latour R, Bagot M, Bensussan A, Socie G, Bouaziz JD. APRIL levels are associated with disease activity in human chronic graft versus host disease. Haematologica. 2016 Mar 18. pii: haematol.2016.145409. [Epub ahead of print]

 

10. de Masson A, Bouaziz JD, Battistella M, Bagot M, Bensussan A. Immunopathology of psoriasis: from bench to bedside. Med Sci (Paris), 2016, 32(3):253-9. doi:10.1051/medsci/20163203009. Epub 2016 Mar 23.

 

11. Battistella M, Janin A, Jean-Louis F, Collomb C, Leboeuf C, Sicard H, Bonnafous C, Dujardin A, Ram-Wolff C, Kadin ME, Bensussan A*, Bagot M*, Michel L*. KIR3DL2 (CD158k) is a potential therapeutic target in primary cutaneous anaplastic large cell lymphoma. Br J Dermatol, 2016 Apr 1. doi: 10.1111/bjd.14626. [Epub ahead of print]

*equal contribution as senior authors

 

12. Brun C, Maginiot F, Cras A, Wong H, Ly Ka So S, Larghero J, Bensussan A, Oddos T, Michel L. Intrinsically aged dermal fibroblasts fail to differentiate into adipogenic lineage. Exp Dermatol, 2016 May 19. doi: 10.1111/exd.13080. [Epub ahead of print]

 

13. De Masson A, Giustiniani J, Marie-Cardine A, Bouaziz JD, Dulphy N, Gossot D, Validire P, Tazi A, Garbar C, Bagot M, Merrouche Y, Bensussan A. Identification of CD245 as myosin 18A, a receptor for surfactant A: A novel pathway for activating human NK lymphocytes. Oncoimmunology. 2016 Jan 13;5(5):e1127493. doi: 10.1080/2162402X.2015.1127493.

 

14. Merrouche Y, Fabre J, Cure H, Garbar C, Fuselier C, Bastid J, Antonicelli F, Al-Daccak R, Bensussan A, Giustiniani J. IL-17E synergizes with EGF and confers in vitro resistance to EGFR-targeted therapies in TNBC cells. Oncotarget. 2016 Jul 23. doi: 10.18632/oncotarget.10804. [Epub ahead of print]

 

15. Vallarelli AF, Rachakonda PS, André J, Heidenreich B, Riffaud L, Bensussan A, Kumar R, Dumaz N. TERT promoter mutations in melanoma render TERT expression dependent on MAPK pathway activation. Oncotarget. 2016 Jul 16. doi: 10.18632/oncotarget.10634. [Epub ahead of print]

 

16. Fabre J, Giustiniani J, Garbar C, Antonicelli F, Merrouche Y, Bensussan A, Bagot M, Al-Dacak R. Targeting the Tumor Microenvironment: The Protumor Effects of IL-17 Related to Cancer Type. Int J Mol Sci. 2016 Aug 30;17(9). pii: E1433. doi: 10.3390/ijms17091433. Review.

 

2015

 

1. de Masson A, Socié G, Bagot M, Bensussan A*, Bouaziz JD. Deficient regulatory B cells in human chronic graft-versus-host disease. Oncoimmunology. 2015 Apr 1;4(7):e1016707. eCollection 2015 Jul.

*corresponding author

 

2. Mombelli S, Cochaud S, Merrouche Y, Garbar C, Antonicelli F, Laprevotte E, Alberici G, Bonnefoy N, Eliaou JF, Bastid J, Bensussan A*, Giustiniani J*. IL-17A and its homologs IL-25/IL-17E recruit the c-RAF/S6 kinase pathway and the generation of pro-oncogenic LMW-E in breast cancer cells. Sci Rep. 2015 Jul 8;5:11874. doi: 10.1038/srep11874.

*equal contribution as senior authors

 

 

3. Bonnefoy N, Bastid J, Alberici G, Bensussan A*, Eliaou JF*. CD39: A complementary target to immune checkpoints to counteract tumor-mediated immunosuppression. Oncoimmunology. 2015 Feb 3;4(5):e1003015. eCollection 2015 May.

*equal contribution as senior authors

 

 

4. Garbar C, Mascaux C, Giustiniani J, Salesse S, Debelle L, Antonicelli F, Merrouche Y, Bensussan A. Autophagy is decreased in triple-negative breast carcinoma involving likely the MUC1-EGFR-NEU1 signalling pathway. Int J Clin Exp Pathol, 2015 May 1;8(5):4344-55. eCollection 2015.

 

5. Laugier F, Finet-Benyair A, André J, Rachakonda PS, Kumar R, Bensussan A, Dumaz N. RICTOR involvement in the PI3K/AKT pathway regulation in melanocytes and melanoma. Oncotarget. 2015 Sep 29;6(29):28120-31. doi: 10.18632/oncotarget.4866.

 

6. Sicard H, Bonnafous C, Morel A, Bagot M, Bensussan A, Marie-Cardine A.

A novel targeted immunotherapy for CTCL is on its way: Anti-KIR3DL2 mAb IPH4102 is potent and safe in non-clinical studies. Oncoimmunology. 2015 Jun 1;4(9):e1022306. eCollection 2015 Sep.

 

7. Engel P, Boumsell L, Balderas R, Bensussan A, Gattei V, Horejsi V, Jin BQ, Malavasi F, Mortari F, Schwartz-Albiez R, Stockinger H, van Zelm MC, Zola H, Clark G. CD Nomenclature 2015: Human Leukocyte Differentiation Antigen Workshops as a Driving Force in Immunology. J Immunol, 2015 Nov 15;195(10):4555-63. doi: 10.4049/jimmunol.1502033.

 

8. Mombelli S, Kwiatkowski F, Abrial C, Wang-Lopez Q, de Boissieu P, Garbar C, Bensussan A, Curé H. Prognostic Factors in Operable Breast Cancer Treated with Neoadjuvant Chemotherapy: Towards a Quantification of Residual Disease. Oncology. 2015 Jan 8. [Epub ahead of print]

 

9. de Masson A, Bouaziz JD, Le Buanec H, Robin M, O'Meara A, Parquet N, Rybojad M, Hau E, Monfort JB, Branchtein M, Michonneau D, Dessirier V, Sicre de Fontbrune F, Bergeron A, Itzykson R, Dhédin N, Bengoufa D, Peffault de Latour R, Xhaard A, Bagot M, Bensussan A*, Socié G*. CD24hiCD27+ and plasmablast-like regulatory B cells in human chronic graft-versus-host disease. Blood. 2015 Jan 20. pii: blood-2014-09-599159. [Epub ahead of print]

*equal contribution as senior authors

 

10. Rahmati M, Petitbarat M, Dubanchet S, Bensussan A, Chaouat G, Ledee N. Colony Stimulating Factors 1, 2, 3 and early pregnancy steps: from bench to bedside. J Reprod Immunol. 2015 Feb 11. pii: S0165-0378(15)00023-6. doi: 10.1016/j.jri.2015.01.005. [Epub ahead of print]

 

11. Schmitt C, Sako N, Bagot M, Bensussan A. Authors' reply. Am J Pathol. 2015 Apr;185(4):1168. doi: 10.1016/j.ajpath.2015.02.001.

 

 

2014

 

1.de Masson A, Guitera P, Brice P, Moulonguet I, Mouly F, Bouaziz JD, Battistella M, Madelaine I, Roux J, Ram-Wolff C, Cayuela JM, Bachelez H, Bensussan A, Michel L, Bagot M. Long-term efficacy and safety of alemtuzumab in advanced primary cutaneous T-cell lymphomas. Br J Dermatol. 2014 Jan 17. doi: 10.1111/bjd.12690. [Epub ahead of print]

 

2. Dulphy N, Henry G, Hemon P, Khaznadar Z, Dombret H, Boissel N, Bensussan A*, Toubert A*.Contribution of CD39 to the immunosuppressive microenvironment of acute myeloid leukaemia at diagnosis. Br J Haematol,  2014 Feb 12. doi: 10.1111/bjh.12774.

*equal contribution as senior authors

 

3. Bouaziz J-D, de Masson A, Bagot M, Bensussan A. IL-10 Producing Regulatory B cells: Where Are We? MOJ Immunol, 2014, 1(1): 00003

 

4. Hu HH, Benfodda M, Dumaz N, Gazal S, Descamps V, Bourillon A, Basset-Seguin N, Riffault A, Ezzedine K, Bagot M, Bensussan A, Saiag P, Grandchamp B, Soufir N. A large french case-control study emphasizes the role of rare Mc1R variants in melanoma risk. Biomed Res Int,2014;2014:925716. doi: 10.1155/2014/925716. Epub 2014 Apr 10.

 

5. Ghazi B, Thonnart N, Bagot M, Bensussan A, Marie-Cardine A. KIR3DL2/CpG ODN Interaction Mediates Sézary Syndrome Malignant T Cell Apoptosis. J Invest Dermatol, 2014 Jul 9. doi: 10.1038/jid.2014.286.

 

6. Sako N, Schiavon V, Bounfour T, Dessirier V, Ortonne N, Olive D, Ram-Wolff C, Michel L, Sicard H, Marie-Cardine A, Bagot M, Bensussan A*, Schmitt C*. Membrane expression of NK receptors CD160 and CD158k contributes to delineate a unique CD4+ T-lymphocyte subset in normal and mycosis fungoides skin. Cytometry A, 2014 Jul 18. doi: 10.1002/cyto.a.22512.

*equal contribution as senior authors

 

7. Bastid J, Bonnefoy N, Eliaou JF, Bensussan A. Lymphocyte-derived interleukin-17A adds another brick in the wall of inflammation-induced breast carcinogenesis. Oncoimmunology.2014 Mar 27;3:e28273. eCollection 2014.

 

8. Moins-Teisserenc H, Daubord M, Clave E, Douay C, Félix J, Marie-Cardine A, Ram-Wolff C, Maki G, Beldjord K, Homyrda L, Michel L, Bensussan A*, Toubert A*, Bagot M*. CD158k Is a Reliable Marker for Diagnosis of Sézary Syndrome and Reveals an Unprecedented Heterogeneity of Circulating Malignant Cells. J Invest Dermatol, 2014 Aug 26. doi: 10.1038/jid.2014.356.

*equal contribution as senior authors

 

9. Bouaziz JD, de Masson A, Le Buanec H, Bagot M, Bensussan A. [Update on regulatory B cells].Med Sci (Paris), 2014 Aug-Sep;30(8-9):721-4. doi: 10.1051/medsci/20143008002.

 

10. Ruocco MG, Chaouat G, Florez L, Bensussan A, Klatzmann D. Regulatory T-cells in pregnancy: historical perspective, state of the art, and burning questions. Front Immunol, 2014 Aug 21;5:389. doi: 10.3389/fimmu.2014.00389. eCollection 2014.

 

11. Sako N, Dessirier V, Bagot M, Bensussan A, Schmitt C.HACE1, a Potential Tumor Suppressor Gene on 6q21, Is Not Involved in Extranodal Natural Killer/T-Cell Lymphoma Pathophysiology.Am J Pathol, 2014 Sep 5. pii: S0002-9440(14)00429-5. doi: 10.1016/j.ajpath.2014.07.011.

 

12. Brun C, Demeaux A, Guaddachi F, Jean-Louis F, Oddos T, Bagot M, Bensussan A, Jauliac S, Michel L. T-Plastin Expression Downstream to the Calcineurin/NFAT Pathway Is Involved in Keratinocyte Migration. PLoS One, 2014 Sep 16;9(9):e104700. doi: 10.1371/journal.pone.0104700. eCollection 2014.

 

13. Brun C, Maginiot F, Cras A, Serrano J, Wong H, Larghero J, Bensussan A, Oddos T, Michel L. Modifications phénotypiques et fonctionnelles des fibroblastes dermiques  primaires liées à l’âge. Ann Dermatol Venereol, 2014 Jun;141(6-7 Suppl 2):S94. doi: 10.1016/j.annder.2014.04.166.

 

14. Rahmati M, Petitbarat M, Dubanchet S, Bensussan A, Chaouat G, Ledee N. Granulocyte-colony stimulating factor related pathways tested on an endometrial ex-vivo model. PLoS One, 2014 Oct 2;9(9):e102286. doi: 10.1371/journal.pone.0102286. eCollection 2014.

 

15. Ibarrola-Villava M, Kumar R, Nagore E, Benfodda M, Guedj M, Gazal S, Hu HH, Guan J, Rachkonda PS, Descamps V, Basset-Seguin N, Bensussan A, Bagot M, Saiag P, Schadendorf D, Martin-Gonzalez M, Mayor M, Grandchamp B, Ribas G, Soufir N. Genes involved in the WNT and vesicular trafficking pathways are associated with melanoma predisposition. Int J Cancer, 2014 Oct 10. doi: 10.1002/ijc.29257.

 

16. Marie-Cardine A, Viaud N, Thonnart N, Joly R, Chanteux S, Gauthier L, Bonnafous C, Rossi B, Bléry M, Paturel C, Bensussan A*, Bagot M*, Sicard H*. IPH4102, a Humanized KIR3DL2 Antibody with Potent Activity against Cutaneous T-cell Lymphoma. Cancer Res, 2014, 74 (21):6060-70. doi: 10.1158/0008-5472.

*equal contribution as senior authors

 

17. Bastid J, Regairaz A, Bonnefoy N, Déjou C, Giustiniani J, Laheurte C, Cochaud S, Laprevotte E, Funck-Brentano E, Hemon P, Gros L, Bec N, Larroque C, Alberici G, Bensussan A*, Eliaou JF*.Inhibition of CD39 Enzymatic Function at the Surface of Tumor Cells Alleviates Their Immunosuppressive Activity. Cancer Immunol Res. 2014 Nov 17. [Epub ahead of print]

2014, 74 (21):6060-70. doi: 10.1158/0008-5472.

*equal contribution as senior authors

 

18. Thonnart N, Caudron A, Legaz I, Bagot M, Bensussan A, Marie-Cardine A. KIR3DL2 is a coinhibitory receptor on Sézary syndrome malignant T cells that promotes resistance to activation-induced cell death. Blood. 2014 Nov 20;124(22):3330-2. doi: 10.1182/blood-2014-09-598995.