Heloisa Cristina Caldas

Hospital de Base

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Mário Abbud‐Filho

Faculdade de Medicina de São José do Rio Preto

Ida Mária Maximina Fernandes-Charpiot

Faculdade de Medicina de São José do Rio Preto

Maria Alice Sperto Ferreira Baptista

Hospital de Base

Fernanda Salomão Gorayeb-Polacchini

Hospital de Base

Irene Fernandes

Universidade de Itaúna

Eny Maria Goloni‐Bertollo

Faculdade de Medicina de São José do Rio Preto

Rosa Sayoko Kawasaki‐Oyama

Faculdade de Medicina de São José do Rio Preto

Glória Elisa Florido Mendes

Faculdade de Medicina de São José do Rio Preto

Domingo Marcolino Braile

Faculdade de Medicina de São José do Rio Preto

ÉC

Érika Cristina Pavarino

Faculdade de Medicina de São José do Rio Preto

Cooperative Institutions

Faculdade de Medicina de São José do Rio Preto

115

John Wiley & Sons (United Kingdom)

Hospital de Base

Universidade de São Paulo

Universidade Estadual Paulista (Unesp)

Hospital de Base

Universidade Estadual de Campinas (UNICAMP)

Universidade Federal de Minas Gerais

Universidade Federal de São Paulo

The Ohio State University

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Activation of HMGB1–TLR4 Pathway and Inflammasome Contribute to Enhanced Inflammatory Response in Extended Criteria and Kidneys With KDPI ≥85%

Transplantation (2019)

Greiciane M. S. Florim Heloisa Cristina Caldas Naiane N Gonçalves Giovanna O.B.E. Bueno Maria Alice Sperto Ferreira Baptista

Background. Metrics for evaluating low-quality kidneys have failed to predict outcomes or reduce the kidney refusal and discard rates. Kidneys from extended-criteria donors (ECDs) and kidneys with ≥85% kidney donor profile indexes (KDPIs) might have different sensitivities to the proinflammatory milieu generated by brain death. We aimed to identify gene expression profile differences in innate immunity pathways between low-quality and ideal kidneys. Methods. Preimplantation kidney biopsies from ECD (n = 41) and standard-criteria donor (n = 39) were evaluated for real-time quantitative polymerase chain reaction gene expression using the TaqMan Gene Expression Array Plates system for genes Toll-like receptor-4 (TLR4), high-mobility group box 1, nuclear factor kappa beta, myeloid differentiation primary response 88, interferon (IFN)-γ, interleukin (IL)1-β, tumor necrosis factor alpha, caspase-1 (CASP1), intercellular adhesion molecule 1, IL-10, heme oxygenase 1 hypoxia-inducible factor 1 (HIF-1), monocyte chemotactic protein 1, transforming growth factor beta 1, TIR-domain containing adapter inducing interferon-β (TRIF), TRIF-related adaptor molecule, interferon regulatory factor 3 (IRF-3), receptor-interacting protein 1, IFNβ-1, and nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin protein 3 complex. Gene expression was also evaluated in kidneys with KDPI ≥85. Results. ECD biopsies showed significantly higher expression of IL-10, TLR4, high-mobility group box 1, IFN-γ, TRIF-related adapter molecule, IRF-3, HIF-1, nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin protein 3 complex, CASP1, and IL-1β ( P < 0.05) compared with standard-criteria donor biopsies. IRF-3, HIF-1, and CASP1 were exclusively upregulated in ECD kidneys. Compared with kidneys with KDPIs <85%, kidneys with KDPIs ≥85% had very similar gene transcripts as those observed in ECD kidneys, except that tumor necrosis factor alpha and monocyte chemotactic protein 1 expression was only elevated in kidneys with KDPIs ≥85%. Significant positive correlations were found between the different genes upregulated and the increase in KDPIs. Conclusions. Our results showed that TLR4 and inflammasome pathways are enhanced in low-quality kidneys and suggest that blocking of some targets might improve transplant outcomes and reduce discard rates.

TRIF

Pyrin domain

HMGB1

Proinflammatory cytokine

10.1097/tp.0000000000003048

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Citations (13)

Sensitivity of human laryngeal squamous cell carcinoma Hep-2 to metrotexate chemoterapy.

PubMed (2012)

Ana Lívia Silva Galbiatti Heloisa Cristina Caldas J Padovani Érika Cristina Pavarino E M Goloni-Bertollo

Methotrexate (MTX) is an antifolate agent that acts inhibiting purine and pyrimidine synthesis. The objective of the study was to evaluate the viability of Hep-2 human laryngeal cancer cells to the treatment with MTX chemotherapy in vitro.Cultured Hep-2 cells were treated with 0.25, 25.0 and 75 μM MTX for 24 h, and their viability was evaluated with Bcl-2-FITC antibody in flow cytometry.The numbers of viable Hep-2 cells after 24 h treatment with 0.25, 25.0 and 75.0 uM MTX were 85.43%, 22.46% and 8.42%, respectively (p < 0.05). Therefore, MTX possesses a dose-dependent effect on viability of Hep-2 cells in vitro.The highest MTX concentration is associated with highest tumor cell sensitivity of human laryngeal cancer cells of Hep-2 line.

Viability assay

Antifolate

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Preimplantation Kidney Biopsies of Extended Criteria Donors Have a Heavier Inflammatory Burden Than Kidneys From Standard Criteria Donors

Transplantation Direct (2017)

Camila M. Mazeti-Felicio Heloisa Cristina Caldas Ida Mária Maximina Fernandes-Charpiot Camila Z. Dezotti Maria Alice Sperto Ferreira Baptista

Donors after brain death develop a systemic proinflammatory state that may predispose the kidneys to injury after transplantation. Because it is not known whether this inflammatory environment similarly affects the kidneys from expanded criteria donor (ECD) and standard criteria donors (SCD), we sought to evaluate differences in the gene expression of inflammatory cytokines in preimplantation biopsies (PIBx) from ECD and SCD kidneys.Cytokines gene expression was measured in 80 PIBx (SCD, 52; ECD, 28) and associated with donor variables.Normal histology and chronic histological lesions were not different between both types of kidneys. ECD kidneys showed significant increase in the transcripts of MCP-1, RANTES, TGF-β1, and IL-10 when compared with SCD. Kidneys presenting normal histology had similar inflammatory profile except by a higher expression of RANTES observed in ECD (P = 0.04). Interstitial fibrosis and tubular atrophy (interstitial fibrosis and tubular atrophy ≥ 1) were associated with higher expression of TGF-β1, RANTES, and IL-10 in ECD compared with SCD kidneys. Cold ischemia time of 24 hours or longer was significantly associated with upregulation of FOXP3, MCP-1, RANTES, and IL10, whereas longer duration of donor hospitalization significantly increased gene expression of all markers. High FOXP3 expression was also associated with lower level of serum creatinine at 1 year. Donor age was not associated with any of the transcripts studied.PIBx of ECD exhibit a higher gene expression of inflammatory cytokines when compared with SCD kidneys. This molecular profile may be a specific ECD kidney response to brain death and may help to predict the posttransplant outcomes of ECD recipients.

Proinflammatory cytokine

Histology

10.1097/txd.0000000000000671

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Citations (6)

P15.06: Distinct FOXP3 Gene Expression in the Peripheral Blood And Inside Renal Allografts Is Associated With Use of MTOR Inhibitors and Extended Criteria Kidney Donor

Transplantation (2022)

Douglas Santos Costa Cinthia Dias Naiane do Nascimento Gonçalves Ida Mária Maximina Fernandes-Charpiot Maria Alice Sperto Ferreira Baptista

Background: Trafficking of regulatory T cells (Tregs) modulate inflammatory response after kidney transplantation. There is scarce information on whether circulating and intragraft Tregs are similarly affected by immunosuppressive drugs (ISD) and by the type of deceased kidney donor. Objectives: 1- to study the simultaneous gene and protein expression of the FOXP3 (forkhead-winged helix transcription factor) in the peripheral blood (PB) and within renal allografts; 2- to correlate FOXP3 expression with the immunosuppressive drugs (ISD) used and kidney donor type. Methods: FOXP3 gene and protein expression were assessed by real-time PCR and immunohistochemical analysis in the peripheral blood (PB) and kidney biopsies (Bx) of patients receiving Tacrolimus (Tac; n=21) or Everolimus (Eve; n=19) at the 3rd month post-Tx. FOXP3 expression was correlated with donor type (standard – SCD or extended criteria – ECD donors), ISD, acute rejection (AR), delayed graft function (DGF), and serum creatinine (sCr) at one year. Results: Eve-treated patients had a longer DGF duration (p=0.04) and a lower frequency of de novo post-Tx diabetes (p=0.03) compared with the Tac group. FOXP3 expression in the PB and Bx was greater in Eve than in Tac-treated patients. Immunohistochemistry did not show differences in the FOXP3 expression for both types of ISD. Recipients of SCD and ECD had similar gene and protein expression inside allografts and in the renal tissue regardless of the ISD. However, in the PB, ECD recipients treated with Eve (ECD/Eve) had higher FOXP3 expression than ECD/Tac (p=0.04) and SCD/Eve (p= 0.01). In the blood Eve and ECD kidney (OR= 5.6; p= 0.04 and OR= 8.7; p= 0,015, respectively) were independently associated with FOXP3 expression while in the Bx only Eve was associate with increased gene expression (OR=5.1; p= 0.03). Conclusion: FOXP3 gene and protein expression does occur differently in the blood and inside allografts from recipients of ECD kidneys treated with mTOR inhibitors. We suggest caution when interpreting studies comparing outcomes based on the measurement of the FOXP3 gene expression in different tissues and compartments and kidney donor types.

Nephrology

10.1097/01.tp.0000889592.24308.e0

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Effect of the COVID-19 Pandemic on Kidney Transplant and on Chronic Dialysis Patients

Transplantation Proceedings (2022)

Thamiris Quiqueto Marinelli Amsei Fernanda Salomão Gorayeb-Polacchini Heloisa Cristina Caldas Ida Mária Maximina Fernandes-Charpiot M.A.S. Ferreira-Baptista

Case fatality rate

Pandemic

10.1016/j.transproceed.2022.02.061

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Citations (3)

Rastreamento de mutações nos genes G6PT1 e AGL em pacientes com glicogenose

Heloisa Cristina Caldas

As doencas de deposito de glicogenio (GSD) incluem aproximadamente 12 tipos, onde existe falta de uma ou de diversas enzimas que metabolizam o glicogenio normal e sao tambem chamadas de glicogenoses. Os acumulos intracelulares do glicogenio afetam principalmente celulas hepaticas, renais, musculo cardiaco e esqueletico. As diferentes formas de GSD foram classificadas de acordo com a ordem cronologica em que os defeitos enzimaticos foram identificados. A GSDI compreende um grupo de doencas raras, com padrao de heranca autossomico recessivo apresentando grande heterogeneidade clinica e bioquimica. Existem dois principais subgrupos, GSDIa e Ib, confirmados em nivel molecular. O subtipo Ia se refere a deficiencia da enzima glicose-6-fosfatase (G6Pase), enzima responsavel pela manutencao da glicose sanguinea; a GSDIb relacionada a deficiencia da glicose-6-fosfato translocase (G6PTl) responsavel pelo transporte da glicose-6-fosfato para o lumen do reticulo endoplasmatico, onde a unidade catalitica da G6Pase esta situada. Outros subgrupos menos comuns foram tambem identificados, tais como o subgrupo Ic que envolve o transporte de fosfato e pirofosfato e, finalmente, Id que envolve a enzima responsavel pelo retomo da glicose do lumen do reticulo para o citosol, existindo, no entanto, controversia com relacao a classificacao desses ultimos subtipos. A GSDID e causada pela deficiencia da enzima bifuncional AGL, uma proteina monomerica com dois sitios cataliticos diferentes: oligo-l, 4-1,4 glucontransferase e amilo-1,6-glucosidase, e para o funcionamento normal, ambas as atividades sao requeridas. A enzima AGL combinada a acao da fosforilase e responsavel pela completa degradacao do glicogenio. Devido a heterogeneidade clinica dos tipos I e m, durante a infância e dificil se diferenciar clinicamente as duas formas, ambas apresentando sintomas similares de hepatomegalia, hipoglicemia, hiperlipidemia,retardo de crescimento entre outros. Neste trabalho foram estudados 14 individuos com sinais clinicos sugestivos de GSDI onde nao foram encontradas alteracoes no gene da G6Pase, responsavel pelo subtipo Ia, esses pacientes sao, portanto classificados como tipo I non-Q. O gene G6PTl foi sequenciado completamente em todos os pacientes estudados e dois deles foram diagnosticados em nivel molecular como sendo na verdade GSDIb. Foram estudados 6 exons do gene AGL, onde estao localizadas a maioria das mutacoes descritas ate o momento no gene, e somente foram detectados polimorfismos em todos os individuos estudados. O protocolo molecular desenvolvido no presente trabalho mostrou-se adequado para o rastreamento de mutacoes, apesar de terem sido diferenciados molecularmente somente dois individuos com subtipo Ib Abstract

10.47749/t/unicamp.2001.222238

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Influence of HLA-A, B, and -DRB1 genes and panel-reactive antibodies on the waitlist time for kidney transplantation in the state of Sao Paulo-Brazil

Transplant Immunology (2024)

Camila Ravazzi-Gauch Ludimila Leite Marzochi Miklos Maximiliano Bajay Marizete Peixoto Medeiros Heloisa Cristina Caldas

Panel reactive antibody

Histocompatibility Testing

Histocompatibility

10.1016/j.trim.2023.101981

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Citations (1)