Transcriptional response of peripheral blood mononuclear cells to recombinant human growth hormone in a routine four-days IGF-I generation test

Welzel, Maik; Appari, Mahesh; Bramswig, Nuria C.; Riepe, Felix G.; Holterhus, Paul‐Martin

doi:10.1016/j.ghir.2011.09.001

Cited by 6 publications

(7 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, rhGH has the potential to regulate all major immune cell types and influence immune function [ 37 ]. The association of rhGH treatment and the immune system we observed in our study corroborates several previous studies on gene expression [ 13 , 14 , 15 ]. Welzel et al performed gene expression studies in PBMC in the frame of the IGFGT and detected inter-individual variability in rhGH-related response.…”

Section: Discussionsupporting

confidence: 93%

“…For the short-term rhGH treatment cohort PBMC isolation from freshly drawn blood samples was performed according to [ 15 ]. Briefly, PBMC were isolated by Ficoll (Biochrom AG, Berlin, Germany) gradient centrifugation, resuspended in RPMI 1640 medium (Gibco, Karlsruhe, Germany) with 10% DMSO, 10% heat-inactivated FCS, penicillin (100 U/mL), streptomycin (100 μg/mL), and L-glutamine (2 mM) and frozen in liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

“…In order to identify in vivo targets of the GH-axis and eventually develop better prediction models to monitor response to rhGH therapy a series of microarray-based gene expression studies has been conducted [ 13 , 14 , 15 ]. Indeed, albeit variable in design and of restricted power due to small sample sizes, transcriptional changes following rhGH treatment were consistently observed in peripheral blood mononuclear cells (PBMCs).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

In vivo Investigations of the Effect of Short- and Long-Term Recombinant Growth Hormone Treatment on DNA-Methylation in Humans

et al. 2015

Self Cite

View full text Add to dashboard Cite

Treatment with recombinant human growth hormone (rhGH) has been consistently reported to induce transcriptional changes in various human tissues including peripheral blood. For other hormones it has been shown that the induction of such transcriptional effects is conferred or at least accompanied by DNA-methylation changes. To analyse effects of short term rhGH treatment on the DNA-methylome we investigated a total of 24 patients at baseline and after 4-day rhGH stimulation. We performed array-based DNA-methylation profiling of paired peripheral blood mononuclear cell samples followed by targeted validation using bisulfite pyrosequencing. Unsupervised analysis of DNA-methylation in this short-term treated cohort revealed clustering according to individuals rather than treatment. Supervised analysis identified 239 CpGs as significantly differentially methylated between baseline and rhGH-stimulated samples (p<0.0001, unadjusted paired t-test), which nevertheless did not retain significance after adjustment for multiple testing. An individualized evaluation strategy led to the identification of 2350 CpG and 3 CpH sites showing methylation differences of at least 10% in more than 2 of the 24 analyzed sample pairs. To investigate the long term effects of rhGH treatment on the DNA-methylome, we analyzed peripheral blood cells from an independent cohort of 36 rhGH treated children born small for gestational age (SGA) as compared to 18 untreated controls. Median treatment interval was 33 months. In line with the groupwise comparison in the short-term treated cohort no differentially methylated targets reached the level of significance in the long-term treated cohort. We identified marked intra-individual responses of DNA-methylation to short-term rhGH treatment. These responses seem to be predominately associated with immunologic functions and show considerable inter-individual heterogeneity. The latter is likely the cause for the lack of a rhGH induced homogeneous DNA-methylation signature after short- and long-term treatment, which nevertheless is well in line with generally assumed safety of rhGH treatment.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

In vivo Investigations of the Effect of Short- and Long-Term Recombinant Growth Hormone Treatment on DNA-Methylation in Humans

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…51]; this study was followed up in a pilot study by Whatmore et al [52], who observed that GH-dependent gene expression could be seen in peripheral blood mononuclear cells from both GHD and TS patients, albeit with less statistical stringency in TS. Since the pilot analysis this phenomenon has been confirmed in adult GHD patients [53], a large data set of normal adults as part of a study on GH doping [54], adult male rats after long-term GH treatment [55], children with GHD and TS after 1 month's treatment with rhGH [41] and, most recently, children with growth disorders undergoing a short-term IGF-1 generation test after 4 days of treatment with GH [56]. …”

Section: Examples Of Pharmacogenomics In Short Stature Disorders: Fromentioning

confidence: 89%

Pharmacogenomics Related to Growth Disorders

et al. 2013

View full text Add to dashboard Cite

Growth disorders resulting in short stature are caused by a wide range of underlying pathophysiological processes. To improve height many of these conditions are treated with recombinant human growth hormone (rhGH). However, substantial inter-individual variability in growth response both in the short and long-term is recognised. Over the last decade, disease-specific growth prediction models have been developed that the clinician can use to define a child's potential response to rhGH and to optimise starting and maintenance doses of rhGH. These models, however, are not able to predict all the variations in treatment response. There has, therefore, been recent interest in using genetic information to contribute to the evaluation of responses to rhGH, including high-throughput technologies for assessing DNA markers (genome) and mRNA transcripts (transcriptome) as pharmacogenomic tools. This review will focus on how these pharmacogenomic approaches are being applied to growth disorders.

show abstract

“…Also, they have been used in the studies of immune dysfunction disorders such as type 2 diabetes, obesity [20,21] and for drug screening to analysis the effect of pharmaceutical agents on corresponding signaling pathways [22][23][24]. PBMCs have been applied for studding the effects of metformin in breast cancer and type 2 diabetes [25], growth hormone treatment [26] and multiple sclerosis [27]. However, PBMCs contain a varied population of immune cells including monocytes, lymphocytes (T Cells, B cells, and NK cells) and dendritic cells.…”

Section: Discussionmentioning

confidence: 99%

Integral membrane protein expression of human CD25 on the cell surface of HEK293 cell line: the available cellular model of CD25 positive to facilitate in vitro developing assays

Dehbashi

Hojati

Motovali-Bashi

et al. 2019

Biomolecular Concepts

View full text Add to dashboard Cite

Typically, CD25 is expressed on the cellular surface of regulatory T (Treg) cells. These cells are significant in regulating the self-tolerance and also preventing the immune system from attacking a person’s own tissues and cells. They promote the cancer progression by playing an important role in evading the immune system. Thus, the experimental procedures was aimed to clone and express human CD25 in HEK293 cell line, as the available cellular model, for the purpose of developing assays to facilitate and enhance the studies on an available CD25 positive cell. The secondary RNA structure of CD25 was evaluated by in silico analysis. Then, cDNA of human CD25 were synthesized from isolated total mRNA of cultured and stimulated PBMCs from blood donors. After cloning the cDNA of CD25 into a pcDNA3.1(+) plasmid, using the effective transfection of the recombinant pcDNA3.1(+) in HEK293, qRT-PCR and flow cytometry methods were used to quantitatively evaluate CD25 transcripts and protein level. There was a 4.8 fold increase in transcripts and a 76.2% increase in protein levels of CD25 when comparing the transfected and control cell lines. The genetically engineered HEK293 cell line expressing Treg cell surface marker of CD25 was introduced in this study for the first time. This cell line can be used to overcome the problematic issues for studying Treg cells including low population of Tregs in peripheral blood, low recovery methods for Treg isolation, time-consuming and non-cost benefit methods in the conditions of in vitro cell culture experiments for the studies focused on the binding of IL-2 to CD25.

show abstract

Transcriptional response of peripheral blood mononuclear cells to recombinant human growth hormone in a routine four-days IGF-I generation test

Cited by 6 publications

References 36 publications

In vivo Investigations of the Effect of Short- and Long-Term Recombinant Growth Hormone Treatment on DNA-Methylation in Humans

In vivo Investigations of the Effect of Short- and Long-Term Recombinant Growth Hormone Treatment on DNA-Methylation in Humans

Pharmacogenomics Related to Growth Disorders

Integral membrane protein expression of human CD25 on the cell surface of HEK293 cell line: the available cellular model of CD25 positive to facilitate in vitro developing assays

Contact Info

Product

Resources

About