João Lacerda Lab

Pathobiology and Treatment of Chronic Graft Versus Host Disease after Hematopoietic Stem Cell Transplantation

This area of research encompasses different subprojects revolving around and emerging from the immunology of hematopoietic stem cell transplantation (HSCT) and how to address chronic graft versus host disease (cGVHD), viral and fungal infections arising in the post-transplant period.

a) Abnormal T-cell homeostasis in cGVHD

This line of research matured from the observation that patients that will develop cGVHD after HSCT have lower numbers of circulating regulatory T cells (Treg) even prior to the emergence of clinical manifestations of the disease. This directed us to prospectively study in detail Treg, Tcon and CD8 homeostasis in patients submitted to unrelated donor HSCT and found that patients developing cGVHD indeed have decreased true naïve and stem cell memory Tcon and Treg, and increased true naïve, SCM and EMRA CD8+ cells early post-transplant. This was the PhD project of Ana Alho MD. These results suggest that these imbalances in the T-cell compartment and that the loss of immunologic tolerance associated with poor Treg recovery are likely associated with the pathobiology of cGVHD.

b) Clinical trials of fresh purified donor Treg in cGVHD

The observation that Treg were decreased in patients with cGVHD led us to design a Phase I/II clinical trial infusing fresh purified Treg from the original HSCT donor into patients with moderate and severe cGVHD. These patients traditionally need to be treated with a combination of immunosuppressive drugs that increase their risk for severe infections. This study was performed under the auspices of the Horizon 2020 EU-funded TREGeneration Project, which brought together several groups in Europe investigating Treg therapies for cGVHD, that we coordinated in Lisbon.  The infusion of increasing numbers of donor Treg proved to be safe and effective, with most patients achieving a partial response of cGVHD and, at the same time, it was possible decrease the dose of the prescribed immunosuppressive drugs.

c) In vitro isolation and expansion of donor versus patient specific Treg

Despite the promising results of the clinical trial with fresh donor Treg, we have been able to select and expand specific Treg after cultures of highly purified Treg with dendritic cells from an HLA-matched sibling. We are currently studying the specificity, potency and mechanisms of suppression of these cells.  This is the PhD project of Carolina Pacini MSc. After this preliminary work we plan to expand the specific Treg under GMP conditions to a clinically significant number and evaluate if they retain their specificity in comparison with the original Treg population. Only then can we envisage utilizing these cells in a new clinical trial.

d) Other projects dissecting the properties of Treg

In a series of more basic projects, we characterized the epigenetic profile of different subtypes of purified human Treg. We discovered that not only the level of methylation of CAMTA gene is able to separate thymic Treg from Tcon, but also that there is a previously unreported strand-bias hemimethylation pattern in FOXP3promoter and TSDR in donors of both genders. In another project we studied the crosstalk between Tcon, Treg and mesenchymal stem cells (MSC). This derived from the observation that patients with acute GVHD treated with MSC had an improvement of the GVHD in parallel with an increase in the peripheral numbers of Treg. We discovered that this interaction with MSC leads to a proliferation of Tcon that acquire a Treg immunophenotype and display a Treg-like epigenetic profile.

CMV and BKV -specific immunity after Hematopoietic Stem Cell Transplantation

In a study performed with the Dana-Farber Cancer Center we have characterized in detail the phenotype and cytokine production of BKV-specific T cells in patients with and without BKV infection after HSCT. This is part of the PhD project of Eduardo Espada MD. More recently, we have been comparing CMV-specific immunity in patients submitted to unrelated HSCT receiving prophylaxis with letermovir versus historic patients treated with the same clinical protocols in the pre-letermovir era, for whom we have biobanked samples. We found that while letermovir effectively prevents the emergence of CMV infection, CMV-specific immunity only develops once letermovir is discontinued.

Genetic susceptibility for CMV and Aspergillus infection after Hematopoietic Stem Cell Transplantation

In the field of infections post-transplant, we have collaborated for many years with Agostinho Carvalho PhD from Universidade do Minho to better understand in both patient and donor the patterns of genetic susceptibility to aspergillus and CMV infections in the post-transplant period.

Dynamic Phenotype of Commercial CAR T cells in Patients with Aggressive Lymphomas

We have been collecting samples from all patients treated with CAR T cells at our institution. In this project, we are interested to characterize immune recovery, CAR T cell fate and persistence in patients treated with Axi-Cell for advanced lymphoma, and to correlate these ancillary results with clinical response to CAR T cells and survival.

Epigenetic crosstalk between DNA and histone modifications in myelodysplastic syndromes and acute myeloid leukaemia

Our overall aim is to study the crosstalk between DNA methylation and histone PTM in MDS and AML. This interplay offers an opportunity to 1) identify new drug targets, 2) discover prognostic biomarkers and 3) study novel mechanisms of resistance to epigenetic drugs. This is the PhD Project of Guilherme Sapinho MD.

Development of Chimeric Antigen Receptor T Cells Targeting Oncological-Associated Chondroitin Sulfate as Cancer Treatment

In this project we aim to develop a novel immunotherapeutic approach based on CAR T cells capable of recognizing oncological-associated forms of chondroitin sulfate. So far, most CAR T cells target aberrantly expressed proteins that, not rarely, are also expressed, at some level, by healthy cells. Our novel approach consists in targeting aberrant post-translational modifications that are more specific to cancer. Indeed, although chondroitin sulfate can be found in multiple tissues of the human body, specific isoforms can be found almost exclusively expressed on different types of cancer. These isoforms arise in cancer cells from aberrant patterns of sulfation in chondroitin sulfate, a glycosaminoglycan commonly found in proteoglycans that are important for the proliferation and invasion properties of cancer cells. Therefore, we hypothesized that a CAR capable of recognizing oncological-associated chondroitin sulfate could be a relevant therapeutic option in the future with strong antitumor responses for multiple types of cancer without employing substantial toxicity over heathy tissues as seen in other CAR T cell therapies. This is the Post-Doctoral Project of Diogo Gomes da Silva PhD.

2013/2016: FCT-Harvard Medical School Portugal Program HMSP-ICT/0001/2011:  Induction of Immune Tolerance in Human Allogeneic Hematopoietic Stem Cell Transplantation. JFLacerda PI

2015/2022: EC Horizon 2020 TREGeneration: Grant Agreement 643776:  Repair of tissue and organ damage in refractory chronic GVHD after HSCT by the infusion of purified allogeneic donor regulatory T lymphocytes. JFLacerda PI

2020/2023: Merck Investigator Studies Program – MISP 2019. CMV infection and CMV-specific T- cell reconstitution in unrelated donor HSCT after T-cell depletion with ATG: prophylaxis with letermovir vs pre-valgancyclovir emptive treatment. JFLacerda PI

2021/2024: FCT- PTDC/MEC- HEM/5281/2020. Generation and in-depth characterization of antigen-specific regulatory T cells to be applied in allogeneic HSCT. JFLacerda PI

2023/2025: Gilead Sciences, PROGRAMA GILEAD GÉNESE 2023, Development of a Chimeric Antigen Receptor T cell therapy against oncologic-specific glycans. Diogo Silva PI

2025/2026: FCT 2023.13874.PEX. Development of a Chimeric Antigen Receptor T cell therapy against oncologic-glycans. Diogo Silva PI

2025/2028: LISBOA2030-FEDER-00678800. Production of donor regulatory T lymphocytes specific to antigens of the patient for the treatment of chronic graft versus host disease after hematopoietic stem cell transplantation. JFLacerda PI

Cunha C, Aversa F, Lacerda JF, Busca A, Kurzai O, Grube M, Löffler M, Maertens JA, Bell AS, Almeida B, Sousa PS, …, Luppi M, Garlanda C, Mantovani A, Velardi A, Romani L, Carvalho A. Genetic deficiency of PTX3 and invasive aspergillosis in stem cell transplantation. New England Journal of Medicine, 370 (5): 421-432, 2014

Alho AC, Kim HT, Chammas MJ, Reynolds CG, Matos TR, Forcade E, Whangbo J, Nikiforow S, Cutler CS, Koreth J, Ho VT, Armand P, Antin JH, Alyea EP, Lacerda JF, Soiffer RJ, Ritz J. Unbalanced recovery of regulatory and effector T cells after allogeneic stem cell transplantation contributes to chronic GVHD. Blood. 127(5):646-657, 2016.

Hirakawa M, Matos T, Liu H, Koreth J, …, Whangbo J, Alho AC, Nikiforow S, Cutler C, Ho VT, Armand P, Alyea EP, Antin JH, Blazar BR, Lacerda JF, Soiffer RJ, Ritz J. Low-dose IL-2 selectively activates subsets of CD4+ Tregs and NK cells. JCI Insight 1(18), 2016

Stappers MHT, Clark AE, Aimanianda V, …, Lacerda JF, Campos A, Carvalho A, Willment JA, Latgé JP, Brown GD. Recognition of DHN-melanin by a C- type lectin receptor is required for immunity to Aspergillus. Nature. 15;555(7696):382-386, 2018

Minskaia E, Saraiva BC, Soares MV, Azevedo RI, Ribeiro RM, Kumar SD, Vieira AIS, Lacerda JF. Molecular markers distinguishing T cell subtypes with TSDR strand-bias methylation. Frontiers in Immunology. 9:2540, 2018

Soares MV, Azevedo RI, Ferreira IA, Bucar SV, Ribeiro AC, Vieira AI, Pereira PN, Ribeiro RM, Ligeiro D, Alho AC, Soares AS, Camacho N, Martins C, Lourenço F, Moreno R, Ritz J, Lacerda JF. Naive and stem cell memory T cell subset recovery reveals opposing reconstitution patterns in CD4 and CD8 T cells in chronic graft versus host disease. Frontiers in Immunology. 10:334, 2019

Azevedo RI, Minskaia E, Fernandes-Platzgummer A, Vieira AIS, Lobato da Silva C, Sampaio Cabral JM, Lacerda JF. Mesenchymal stem cells induce a functional Treg- like population, but do not expand natural Treg in vitro. Stem Cells. 38(8):1007-1019, 2020

Espada E, Cheng MP, Kim HT, …, Soares MVD, Lacerda JF, …, Soiffer RJ, Marty FM, Ritz JR. BK vírus-specific T-cell immune reconstitution after allogeneic hematopoietic cell transplantation. Blood Advances 4(9):1881-1893, 2020

Soares MVD, Gomez VE, Azevedo RI, Pereira PNG, Velázquez TC, Garcia-Calderón CB, Tharmaratnam K, Cabral IA, Ribeiro AC, Mendes L, Juncal C, Roncon S, Pais AT, Alho AC, …, Ritz J, Vaz CP, Perez-Simon JA, Lacerda JF. Phase I/II Clinical Trials of Donor-Derived Puried Regulatory T Cells for the Treatment of Steroid-Refractory Chronic Graft Versus Host Disease. Blood 140 (Supplement 1): 880–882, 2022. (American Society of Hematology Oral Presentation)

Doglio M, Crossland RE, Alho AC, Penack O, Dickinson AM, Stary G, Lacerda JF, Eissner G, Inngjerdingen M. Cell-based therapy in prophylaxis and treatment of chronic graft-versus-host disease. Frontiers in Immunology 13, 1045168, 2023

Pacini CP, Soares MVD, Lacerda JF. The impact of regulatory T cells on the graft-versus-leukemia effect. Frontiers in Immunology 15, 1339318, 2024

Find a complete list of publications here.