Elevated level of HSPA1L mRNA correlates with graft-versus-host disease
Introduction
Allogeneic-HSCT is used to cure numerous malignant and non-malignant haematological disorders [43]. More than 25,000 allo-HSCTs are performed annually world-wide [33]. However, GVHD still remains a major risk factor for allo-HSCT; directly contributing to a transplant related mortality rate of 26% for matched sibling and 35% for matched unrelated donor transplants [34]. Despite all the improvements achieved to date with respect to treatment and patient after-care, there are a limited number of validated biomarkers for GVHD, in particular for cGVHD [23], [32].
Heat Shock Proteins (HSPs) are highly conserved and ubiquitously present in all organisms [50]. Various physiological roles have been attributed to circulating HSPs such as (i) molecular chaperones [14], (ii) down-regulation of autoimmune responses [3], (iii) down-regulation of T cell responses [47] and (iv) cytoprotection [25], [27]. As molecular chaperones, they play a role in protein folding, repair and maintenance of cellular homeostasis [14], [50]. Under normal conditions HSPs comprise only 5% of the total intracellular proteins; however, under stress, the level elevates to approximately 15% [45]. Heat Shock Protein 70 family (HSPA) is induced by different stimuli such as: heat [41], metal ions and oxidative stress [51], infection [30], inflammation [40] and anti-cancer drugs [24]. Investigations have exhibited HSPs on the surface of cells [28], in sera [37] and in secretions from viable cells [19].
Heat Shock Proteins are characterised according to their molecular weight and their role has been investigated in a variety of diseases [29], [52]. The HSPA family comprises of thirteen members [10]; two members in this investigation belong to the heat inducible group; HSPA1A (HSP70-1) and HSPA1B (HSP70-2), and the third constitutively expressed member; HSPA1L (HSP70-hom). The two heat inducible members are collectively known as HSP0i [22]. The exons of HSPA1A and HSPA1B vary by only six single bases [44], which generates a difference of two amino-acids in the protein sequences [22].
Heat Shock Proteins have been shown to be associated with responses to inflammation and allograft rejection [20], [48]. HSPAs have also exhibited crucial functions during stress and in prediction of allo-HSCT outcome [12]. HSPA1B protein has been shown to be correlated to severity of graft-versus-host reactivity (GVHR) in the in vitro skin explant model [20].
Section snippets
Objective
The overall objective of this study was to test whether there was a significant differential HSPA1L, HSPA1B and/or HSPA1A mRNA expression level in clinical skin biopsies and whole blood from patients with or without aGVHD and cGVHD that could predict disease incidence and/or severity prior to GVHD onset.
Ethics and consent
The study was approved by the Newcastle and North Tyneside Research Ethics Committee I, and all patients and normal volunteers gave their written informed consent for the collection of both skin and whole blood specimens as well as for molecular testing. The investigations were conducted in accordance with the Helsinki Declaration, and consents were taken prospectively by trained personnel. Both aGVHD and cGVHD were assessed by clinicians according to the NIH consensus. All the clinical data
Patients with histopathological skin aGVHD grades II–III had significantly lower HSPA1L mRNA levels
Quantitative RT-PCR was used to investigate HSPA1L, HSPA1B and HSPA1A mRNA expression levels in clinical skin biopsies obtained from allo-HSCT patients with and without aGVHD at various time-points post-transplant at the onset of aGVHD. The histopathological grading was used to analyse the HSPA expressions. Interestingly, HSPA1L expression was significantly lower in patients with grade II–III aGVHD when compared to those without aGVHD (p ≤ 0.05) and normal controls (p ≤ 0.01). As expected, there
Discussion
Even though the field of allo-HSCT has advanced progressively, GVHD is still a major post-transplant complication. There are numerous risk factors that predispose patients to GVHD. However, the incidence of prior aGVHD can increase the likelihood to cGVHD [4], [5], [7]. In this investigation we have looked at both acute and cGVHD patients with regard to HSPA1L, HSPA1B and HSPA1A expression in clinical skin biopsies and in whole blood samples. The potential role of HSP0i in initiation and GVHD
Funding
This investigation was supported by the European Commission Marie Curie Research Training Network MCRTN-TRANSNET (Contract no. 512253), FP6 EC Stemdiagnostics (Contract no. 037703) and the FP7 Marie Curie Initial Training Network CELLEurope (Contract No: 315963).
Disclosure
The authors declare no conflicts of interest.
Author's contributions
SA: performed the research and data analysis and wrote the manuscript.
BT, JN, SA, KBK, XNW and AD: designed the study and commented on the manuscript.
AD, JN and KFP: contributed to data interpretation.
KFP: contributed to statistical analysis.
MC: contributed clinical samples for the study.
Acknowledgements
The authors thank the patients who have donated samples for this study. The authors also thank E. Douglas and L.J. Ambler for their technical support.
References (52)
- et al.
Chronic GVHD risk score: a Center for International Blood and Marrow Transplant Research analysis
Blood
(2011) - et al.
Risk factors for chronic graft-versus-host disease after HLA-identical sibling bone marrow transplantation
Blood
(1990) - et al.
The heat shock protein 70 family: highly homologous proteins with overlapping and distinct functions
FEBS Lett
(2007) - et al.
National Institutes of Health consensus development project on criteria for clinical trials in chronic graft-versus-host disease: I. Diagnosis and staging working group report
Biol Blood Marrow Transplant
(2005) - et al.
Reduced levels of Hsp70 result in a therapeutic effect of 15-deoxyspergualin on acute graft-versus-host disease in (DA × LEW)F1 rats
Immunobiology
(2000) - et al.
Hsp70 release from peripheral blood mononuclear cells
Biochem Biophys Res Commun
(2004) - et al.
Guidelines for the nomenclature of the human heat shock proteins
Cell Stress Chaperones
(2009) - et al.
Clinical applications for biomarkers of acute and chronic graft-versus-host disease
Biol Blood Marrow Transplant
(2012) - et al.
Anticancer drugs cause release of exosomes with heat shock proteins from human hepatocellular carcinoma cells that elicit effective natural killer cell antitumor responses in vitro
J Biol Chem
(2012) Discovery and validation of graft-versus-host disease biomarkers
Blood
(2013)
Heat shock proteins as regulators of the immune response
Lancet
Heat shock proteins
Journal of Biological Chemistry
Inducible heat shock protein 70 reduces T cell responses and stimulatory capacity of monocyte-derived dendritic cells
Journal of Biological Chemistry
Circulating heat shock protein 70 and progression in patients with chronic myeloid leukemia
Leuk Res
Metal ions induced heat shock protein response by elevating superoxide anion level in HeLa cells transformed by HSE-SEAP reporter gene
Toxicology
Heat shock factors: integrators of cell stress, development and lifespan
Nat Rev Mol Cell Biol
The functions of animal microRNAs
Nature
Activation of T cells recognizing self 60-kD heat shock protein can protect against experimental arthritis
J Exp Med
Impact of prior acute GVHD on chronic GVHD outcomes: a chronic graft versus host disease consortium study
Leukemia
MicroRNAs: the missing link in the biology of graft-versus-host disease?
Front Immunol
The microRNA.org resource: targets and expression
Nucleic Acids Res
HSP70-hom gene polymorphism in allogeneic hematopoietic stem-cell transplant recipients correlates with the development of acute graft-versus-host disease
Transplantation
hsp70 genes in the human genome: conservation and differentiation patterns predict a wide array of overlapping and specialized functions
BMC Evol Biol
The potential role of HSP70 in predicting outcome in haematopoietic stem cell transplants — genomic and functional studies
Role of the major heat shock proteins as molecular chaperones
Annu Rev Cell Biol
Clinical manifestations of graft-versus-host disease in human recipients of marrow from HL-A-matched sibling donors
Transplantation
Cited by (4)
Heat Shock 70 Protein Genes and Genetic Susceptibility to Apical Periodontitis
2016, Journal of EndodonticsCitation Excerpt :In contrast, it has been suggested that HSPA1L does not influence the risk to infectious morbidities or specific organ failure or increased mortality risk in surgical care unit patients (33). Also, the presence of this protein has been suggested as an early predictor for chronic graft-versus-host disease at the mRNA expression level (34). HSPA6 is expressed in basal and stress-inducible human keratinocytes and is essential to increasing survival of cells exposed to increased temperatures and chemicals; it was also shown to provide critical benefits to buffering neuronal migration from cellular stress in the human brain (35).
MAPKAP kinase 2–mediated phosphorylation of HspA1L protects male germ cells from heat stress–induced apoptosis
2019, Cell Stress and Chaperones