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1.
Exp Hematol ; 67: 41-48, 2018 11.
Article in English | MEDLINE | ID: mdl-30195076

ABSTRACT

Ponatinib represents a remarkable progress in the treatment of heavily pretreated chronic myelogenous leukemia (CML) and de novo Philadelphia chromosome-positive ALL patients despite significant toxicity in clinical trials. To date, "real-life" data remain few and the use of ponatinib in this setting and its consequences remain mostly unknown. We report, within a national observational study, the use of ponatinib in unselected CML patients who had previously failed ≥2 lines of tyrosine kinase inhibitor (TKI) therapy (or one line if an Abelson (ABL)T315I mutation was identified), in real-life conditions (2013-2014) in a compassionate program. Our analysis has been focused on 48 chronic phase CML patients recorded. With a median follow-up of 26.5 months since ponatinib initiation, the overall survival (OS) rates (80.5% at 3 years) and cumulative incidence of major molecular response (81.8% at 18 months) were similar to those of the phase II study, with no influence of BCR-ABL mutations nor the reason of ponatinib prescription. A specific subanalysis of the preexisting cardiovascular risk factors and events occurring on ponatinib is described. These events occurred after a median time on ponatinib of 5.8 months (excluding hypertension) and were observed in 29/48 patients (47%), even in those already on anti-aggregants/coagulants. The majority were not severe and resolved, but two cases were fatal. Other hematological or nonhematological nonvascular adverse events were similar to those previously described in trials. This observational study reports similar rates of survival, molecular responses, and a slight increase in the cardiovascular toxicity of ponatinib in real-life conditions, prompting improved control of cardiovascular risk factors and selection of patients.


Subject(s)
Antineoplastic Agents/therapeutic use , Imidazoles/therapeutic use , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Protein Kinase Inhibitors/therapeutic use , Pyridazines/therapeutic use , Salvage Therapy , Adolescent , Adult , Aged , Aged, 80 and over , Antineoplastic Agents/adverse effects , Cardiovascular Diseases/chemically induced , Compassionate Use Trials , Drug Resistance, Neoplasm , Female , Genes, abl , Humans , Imidazoles/adverse effects , Intention to Treat Analysis , Kaplan-Meier Estimate , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/enzymology , Male , Middle Aged , Patient Selection , Pragmatic Clinical Trials as Topic , Protein Kinase Inhibitors/adverse effects , Pyridazines/adverse effects , Survival Analysis , Treatment Failure , Young Adult
2.
J Pathol ; 243(1): 51-64, 2017 09.
Article in English | MEDLINE | ID: mdl-28603917

ABSTRACT

Castration-resistant prostate cancer is a lethal disease. The cell type(s) that survive androgen deprivation remain poorly described, despite global efforts to understand the various mechanisms of therapy resistance. We recently identified in wild-type (WT) mouse prostates a rare population of luminal progenitor cells that we called LSCmed according to their FACS profile (Lin- /Sca-1+ /CD49fmed ). Here, we investigated the prevalence and castration resistance of LSCmed in various mouse models of prostate tumourigenesis (Pb-PRL, Ptenpc-/- , and Hi-Myc mice). LSCmed prevalence is low (∼8%, similar to WT) in Hi-Myc mice, where prostatic androgen receptor signalling is unaltered, but is significantly higher in the two other models, where androgen receptor signalling is decreased, rising up to more than 80% in Ptenpc-/- prostates. LSCmed tolerate androgen deprivation and persist or are enriched 2-3 weeks after castration. The tumour-initiating properties of LSCmed from Ptenpc-/- mice were demonstrated by regeneration of tumours in vivo. Transcriptomic analysis revealed that LSCmed represent a unique cell entity as their gene expression profile is different from luminal and basal/stem cells, but shares markers of each. Their intrinsic androgen signalling is markedly decreased, explaining why LSCmed tolerate androgen deprivation. This also illuminates why Ptenpc-/- tumours are castration-resistant since LSCmed represent the most prevalent cell type in this model. We validated CK4 as a specific marker for LSCmed on sorted cells and prostate tissues by immunostaining, allowing for the detection of LSCmed in various mouse prostate specimens. In castrated Ptenpc-/- prostates, there was significant proliferation of CK4+ cells, further demonstrating their key role in castration-resistant prostate cancer progression. Taken together, this study identifies LSCmed as a probable source of prostate cancer relapse after androgen deprivation and as a new therapeutic target for the prevention of castrate-resistant prostate cancer. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Subject(s)
Biomarkers, Tumor/deficiency , Cell Proliferation , Neoplastic Stem Cells/enzymology , PTEN Phosphohydrolase/deficiency , Prostatic Neoplasms, Castration-Resistant/enzymology , Androgen Antagonists/pharmacology , Animals , Antineoplastic Agents, Hormonal/pharmacology , Ataxin-1/metabolism , Biomarkers, Tumor/genetics , Cell Lineage , Cell Proliferation/drug effects , Drug Resistance, Neoplasm , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Genetic Predisposition to Disease , Integrin alpha6/metabolism , Keratin-4/metabolism , Male , Mice, Inbred C57BL , Mice, Knockout , Neoplasm Recurrence, Local , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/pathology , Neoplastic Stem Cells/transplantation , Oligonucleotide Array Sequence Analysis , PTEN Phosphohydrolase/genetics , Phenotype , Prostatic Neoplasms, Castration-Resistant/drug therapy , Prostatic Neoplasms, Castration-Resistant/genetics , Prostatic Neoplasms, Castration-Resistant/pathology , Receptors, Androgen/drug effects , Receptors, Androgen/metabolism , Signal Transduction
3.
Gen Comp Endocrinol ; 220: 133-42, 2015 Sep 01.
Article in English | MEDLINE | ID: mdl-25888939

ABSTRACT

Local overexpression of prolactin (PRL) in the prostate of Pb-PRL transgenic mice induces benign prostate tumors exhibiting marked amplification of the epithelial basal/stem cell compartment. However, PRL-activated intracellular signaling seems to be restricted to luminal cells, suggesting that basal/stem cells may not be direct targets of PRL. Given their described role as prostate cancer-initiating cells, it is important to understand the mechanisms that regulate basal/stem cells. In this study, we evaluated whether PRL can act directly on these cells, by growing them as prostaspheres. For this, primary 3D prostasphere cultures were prepared from unfractionated cells isolated from freshly harvested human and mouse benign prostate tissues and subjected to PRL stimulation in vitro. None of the various concentrations of PRL tested showed any effects on the sizes or numbers of the prostaspheres generated. In addition, neither activation of canonical PRL-induced signaling pathways (Stat5, Stat3 or Erk1/2) nor increased expression of the proliferation marker Ki-67 were detected by immunostaining in PRL-stimulated prostaspheres. Consistent with the absence of response, PRL receptor mRNA levels were generally undetectable in mouse sphere cells. We conclude that human and mouse prostate basal/stem cells are not direct targets of PRL action. The observed amplification of basal/stem cells in Pb-PRL prostates might be due to paracrine mechanisms originating from PRL action on other cell compartments. Our current efforts are aimed at unraveling these mechanisms.


Subject(s)
Prolactin/metabolism , Prostatic Neoplasms/metabolism , Stem Cells/metabolism , Animals , Humans , Male , Mice , Receptors, Prolactin , Signal Transduction
4.
Mol Endocrinol ; 29(5): 667-81, 2015 May.
Article in English | MEDLINE | ID: mdl-25793405

ABSTRACT

Adult stem/progenitor cells are found in many tissues, where their primary role is to maintain homeostasis. Recent studies have evaluated the regulation of adult stem/progenitor cells by prolactin in various target tissues or cell types, including the mammary gland, the prostate, the brain, the bone marrow, the hair follicle, and colon cancer cells. Depending on the tissue, prolactin can either maintain stem cell quiescence or, in contrast, promote stem/progenitor cell expansion and push their progeny towards differentiation. In many instances, whether these effects are direct or involve paracrine regulators remains debated. This minireview aims to overview the current knowledge in the field.


Subject(s)
Adult Stem Cells/physiology , Prolactin/physiology , Animals , Cell Differentiation , Cell Self Renewal , Humans , Signal Transduction
5.
Adv Exp Med Biol ; 846: 221-42, 2015.
Article in English | MEDLINE | ID: mdl-25472541

ABSTRACT

The physiological role of prolactin (PRL) in the prostate gland is not clearly understood. Genetically-modified mouse models that have invalidated actors of the PRL signaling axis failed to identify an essential regulatory function on this tissue. However, a large body of evidence suggests an important role for PRL in prostate tumorigenesis. Mainly through the activation of its downstream target STAT5, PRL can induce growth and survival of prostate cancer cells and tissues in several experimental settings. In the clinic, PRL expression and STAT5 activation in human prostate tumors correlate with disease severity. Available data point to a role of local (autocrine/paracrine) rather than circulating (endocrine) PRL in the induction of disease progression. In mice, transgenic expression of PRL in the prostate leads to enhanced epithelial hyperplasia and dysplasia, with amplification of basal/stem cells which have been recently identified as prostate cancer-initiating cells. Thus, targeting PRL receptor (PRLR)/STAT5 signaling may provide an alternative therapy for the treatment of prostate cancer. Corresponding targeted therapies currently in preclinical development include antagonists or blocking antibodies for the PRLR and small molecule inhibitors directed against the tyrosine kinase JAK2 upstream of STAT5. Present efforts are aimed at validating these therapies for the treatment of prostate cancer, while understanding the mechanisms of disease progression induced by PRL/STAT5.


Subject(s)
Carcinogenesis , Prolactin/physiology , Prostate/pathology , Prostatic Neoplasms/pathology , Animals , Humans , Male , Mice , Prolactin/pharmacology , Prostate/drug effects , Prostatic Neoplasms/therapy , Receptors, Androgen/physiology , STAT5 Transcription Factor/physiology
6.
Am J Pathol ; 184(11): 3105-19, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25193592

ABSTRACT

Current androgen ablation therapies for prostate cancer are initially successful, but the frequent development of castration resistance urges the generation of alternative therapies and represents an important health concern. Prolactin/signal transducer and activator of transcription 5 (STAT5) signaling is emerging as a putative target for alternative treatment for prostate cancer. However, mechanistic data for its role in development or progression of prostate tumors are scarce. In vivo mouse studies found that local prolactin induced the amplification of prostate epithelial basal/stem cells. Because these cells are proposed cells of origin for prostate cancer and disease recurrence, we looked further into this amplification. Our results indicated that sustained Stat5 activation was associated with the occurrence of abnormal basal/stem cell clusters in prostate epithelium of prostate-specific prolactin-transgenic mice. Analysis of epithelial areas containing these clusters found high proliferation, Stat5 activation, and expression of stem cell antigen 1. Furthermore, enhanced prolactin signaling also led to amplification of a luminal cell population that was positive for stem cell antigen 1. These cells may originate from amplified basal/stem cells and might represent important progenitors for tumor development in prostate epithelium. These data provide a deeper understanding of the initial stages of prostate tumorigenesis induced by prolactin to help determine whether this hormone or its downstream messengers could be useful targets for prostate cancer treatment in the future.


Subject(s)
Carcinogenesis/metabolism , Prolactin/metabolism , Prostate/metabolism , STAT5 Transcription Factor/metabolism , Signal Transduction/physiology , Animals , Carcinogenesis/pathology , Cell Differentiation , Cell Proliferation , Male , Mice , Mice, Transgenic , Neoplastic Stem Cells , Prolactin/genetics , Prostate/pathology
7.
Int J Pediatr Endocrinol ; 2014(1): 9, 2014.
Article in English | MEDLINE | ID: mdl-24949022

ABSTRACT

BACKGROUND: Early diagnosis of initial metabolic derangements in young obese children could influence their management; however, this impairment is frequently not overt, but subtle and undetectable by routinely used clinical assays. Our aim was to evaluate the ability of serum proteomic analysis to detect these incipient metabolic alterations in comparison to standard clinical methods and to identify new candidate biomarkers. METHODS: A cross-sectional study of fasting serum samples from twenty-two prepubertal, Caucasian obese (OB; 9.22 ± 1.93 years; 3.43 ± 1.08 BMI-SDS) and twenty-one lean controls (C; 8.50 ± 1.98 years; -0.48 ± 0.81 BMI-SDS) and a prospective study of fasting serum samples from twenty prepubertal, Caucasian obese children (11 insulin resistant [IR]) before (4.77 ± 1.30 BMI-SDS) and after weight reduction (2.57 ± 1.29 BMI-SDS) by conservative treatment in a reference hospital (Pros-OB) was performed. Proteomic analysis (two-dimension-eletrophoresis + mass spectrometry analysis) of serum and comparative evaluation of the sensitivity of routinely used assays in the clinics to detect the observed differences in protein expression level, as well as their relationship with anthropometric features, insulin resistance indexes, lipid profile and adipokine levels were carried out. RESULTS: Study of the intensity data from proteomic analysis showed a decrease of several isoforms of apolipoprotein-A1, apo-J/clusterin, vitamin D binding protein, transthyretin in OB vs. C, with some changes in these proteins being enhanced by IR and partially reversed after weight loss. Expression of low molecular weight isoforms of haptoglobin was increased in OB, enhanced in IR and again decreased after weight loss, being positively correlated with serum interleukin-6 and NAMPT/visfatin levels. After statistical correction for multiple comparisons, significance remained for a single isoform of low MW haptoglobin (OB vs. C and IR vs. non-IR) and Apo A1 (IR vs. non-IR). Assays routinely used in the clinical setting (ELISA/kinetic nephelometry), only partially confirmed the changes observed by proteomic analysis (ApoA1 and haptoglobin). CONCLUSION: Proteomic analysis can allow for the identification of potential new candidate biomarkers as a complement to routinely used assays to detect initial changes in serum markers of inflammation and lipid metabolism impairment in young obese children.

8.
J Gerontol A Biol Sci Med Sci ; 69(1): 34-43, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23873966

ABSTRACT

Growth hormone receptor-null (GHR(-/-)) mice are dwarf, insulin sensitive, and long-lived in spite of increased adiposity. However, their adiposity is not uniform, with select white adipose tissue (WAT) depots enlarged. To study WAT depot-specific effects on insulin sensitivity and life span, we analyzed individual WAT depots of 12- and 24-month-old GHR(-) (/-) and wild-type (WT) mice, as well as their plasma levels of selected hormones. Adipocyte sizes and plasma insulin, leptin, and adiponectin levels decreased with age in both GHR(-) (/-) and WT mice. Two-dimensional gel electrophoresis proteomes of WAT depots were similar among groups, but several proteins involved in endocytosis and/or cytoskeletal organization (Ehd2, S100A10, actin), anticoagulation (S100A10, annexin A5), and age-related conditions (alpha2-macroglobulin, apolipoprotein A-I, transthyretin) showed significant differences between genotypes. Because Ehd2 may regulate endocytosis of Glut4, we measured Glut4 levels in the WAT depots of GHR(-) (/-) and WT mice. Inguinal WAT of 12-month-old GHR(-) (/-) mice displayed lower levels of Glut4 than WT. Overall, the protein changes detected in this study offer new insights into possible mechanisms contributing to enhanced insulin sensitivity and extended life span in GHR(-) (/-) mice.


Subject(s)
Adipocytes/metabolism , Adipose Tissue, White/metabolism , Aging/physiology , DNA/genetics , Gene Expression Regulation, Developmental , Insulin Resistance , Receptors, Somatotropin/genetics , Adipocytes/cytology , Animals , Blotting, Western , Disease Models, Animal , Electrophoresis, Gel, Two-Dimensional , Male , Mice , Mice, Inbred C57BL , Obesity/genetics , Obesity/metabolism , Receptors, Somatotropin/metabolism
9.
Proteomics ; 13(3-4): 674-85, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23019135

ABSTRACT

Growth hormone (GH) is a protein secreted by the anterior pituitary and circulates throughout the body to exert important actions on growth and metabolism. GH stimulates the secretion of insulin-like growth factor-I (IGF-I) that mediates some of the growth promoting actions of GH. The GH/IGF-I axis has recently been recognized as important in terms of longevity in organisms ranging from Caenorhabditis elegans to mice. For example, GH transgenic mice possess short lifespans while GH receptor null (GHR-/-) mice have extended longevity. Thus, the actions of GH (or IGF-I) or lack thereof impact the aging process. In this review, we summarize the proteomic analyses of plasma and white adipose tissue in these two mouse models of GH action, i.e. GH transgenic and GHR-/- mice. At the protein level, we wanted to establish novel plasma biomarkers of GH action as a function of age and to determine differences in adipose tissue depots. We have shown that these proteomic approaches have not only confirmed several known physiological actions of GH, but also resulted in novel protein biomarkers and targets that may be indicative of the aging process and/or new functions of GH. These results may generate new directions for GH and/or aging research.


Subject(s)
Aging/metabolism , Growth Hormone/physiology , Proteome/metabolism , Adipose Tissue, White/metabolism , Animals , Blood Proteins/metabolism , Gene Knockout Techniques , Humans , Insulin/physiology , Insulin Resistance , Insulin-Like Growth Factor I/physiology , Lipid Metabolism , Proteomics , Receptors, Somatotropin/deficiency , Receptors, Somatotropin/genetics
10.
Metabolism ; 61(11): 1646-57, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22658937

ABSTRACT

UNLABELLED: In most obese patients there is an inflammatory state characterized by lipid abnormalities, hyperleptinemia and hyperinsulinemia. OBJECTIVE: The objective was to identify mechanisms involved in leptin's role in the attenuation of the response to insulin using a proteomic approach. MATERIAL/METHODS: We studied the serum proteomic profile of rats treated by central leptin infusion followed by an injection of insulin. We analyzed the relationship between these proteins and serum cytokine and apolipoprotein levels. RESULTS: Out of 81 protein spots, intensity differences were found in 11, corresponding to 5 proteins: three isoforms of α1 macroglobulin; three of haptoglobin and serum amyloid P component-precursor. All of these are acute-phase proteins involved in inflammation and are correlated with cytokine levels. Additionally, two apolipoprotein E and two apolipoprotein A1 isoforms were identified and were found to correlate with LDL and HDL. CONCLUSIONS: Our results indicate that increased leptin and insulin levels change these circulating proteins, thus promoting systemic inflammation and changing lipid metabolism.


Subject(s)
Cytokines/blood , Insulin/administration & dosage , Leptin/administration & dosage , Lipoproteins/blood , Animals , Body Weight , Electrophoresis, Gel, Two-Dimensional , Enzyme-Linked Immunosorbent Assay , Leptin/genetics , Male , RNA, Messenger/genetics , Rats , Rats, Wistar , Real-Time Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
11.
Age (Dordr) ; 34(5): 1225-37, 2012 Oct.
Article in English | MEDLINE | ID: mdl-21953241

ABSTRACT

Unintentional weight loss (wasting) in the elderly is a major health concern as it leads to increased mortality. Several studies have focused on muscle loss, but little is known about the mechanisms giving rise to loss of fat mass at old ages. To investigate potential mechanisms, white adipose tissue (WAT) characteristics and proteomic profiles were compared between adult (10-12-month-old) and aged (22-24-month-old) wild-type mice. Four individual WAT depots were analyzed to account for possible depot-specific differences. Proteomic profiles of WAT depots, along with body weights and compositions, plasma levels of insulin, leptin and adiponectin, insulin tolerance, adipocyte sizes, and products of oxidative damage in each WAT depot were determined. We found that lean mass remained constant while fat mass and insulin tolerance were decreased in old age, as were adipocyte sizes in the WAT depots. Proteomic results showed increased levels of enolase, pyruvate dehydrogenase E1ß, NAD(+)-dependent isocitrate dehydrogenase α, and ATP synthase subunit ß, and decreased levels of carbonic anhydrase 3 in WAT of aged mice. These data suggest increased aerobic glucose oxidation in wasting WAT, consistent with decreased insulin signaling. Also, Cu/Zn superoxide dismutase and two chaperones were increased in aged WAT depots, indicating higher stress resistance. In agreement, lipid peroxidation (HNE-His adducts) increased in old age, although protein oxidation (carbonyl groups) showed no increase. In conclusion, features of wasting WAT were similar in the four depots, including decreased adipocyte sizes and alterations in protein expression profiles that indicated decreased insulin sensitivity and increased lipid peroxidation.


Subject(s)
Adipose Tissue, White/metabolism , Adipose Tissue/metabolism , Aging/physiology , Insulin Resistance/physiology , Insulin/metabolism , Obesity/metabolism , Oxidative Stress , Adipocytes/metabolism , Adipose Tissue, White/pathology , Animals , Disease Models, Animal , Male , Mice , Mice, Inbred C57BL , Obesity/pathology , Proteomics
12.
Obesity (Silver Spring) ; 20(1): 101-11, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21779095

ABSTRACT

The widespread prevalence of obesity has lead to extensive research on white adipose tissue (WAT), which frequently uses the C57BL/6J mouse strain as a model. In many studies, results obtained in one WAT depot are often extrapolated to all WAT. However, functional differences among WAT depots are now becoming apparent. Thus, to identify the molecular mechanisms responsible for WAT depot-specific differences under "normal" conditions, four C57BL/6J mouse WAT depots (inguinal, mesenteric, epididymal, and retroperitoneal) were analyzed. Depot proteomic profiles, along with weights, protein contents, adipocyte sizes and oxidative stress were determined. Mesenteric WAT had almost twice the protein content of the other depots analyzed. Mean adipocyte size was highest in epididymal and lowest in mesenteric and inguinal depots. The proteome of inguinal WAT displayed low levels of enzymes involved in ATP generation, glucose and lipid metabolism, and antioxidant proteins. Higher levels of these proteins were observed in mesenteric and epididymal WAT, with variable levels in the retroperitoneal depot. Some of these proteins showed depot-specific correlations with plasma levels of insulin, leptin, and adiponectin. In agreement with the proteomic data, levels of the antioxidant protein heat shock protein ß1 (HSPß1) also were lower in inguinal WAT when analyzed by western blotting and immunohistochemistry. Also, lipid peroxidation products showed similar trends. Our results are consistent with lower triglyceride turnover and lower oxidative stress in inguinal than mesenteric and epididymal WAT. The observed WAT depot-specific differences provide clues as to the mechanisms leading to these depots' respective diverse functions.


Subject(s)
Adipocytes/pathology , Adipose Tissue, White/pathology , Lipid Peroxidation , Obesity/pathology , Animals , Blotting, Western , Gene Expression , Immunohistochemistry , Lipid Peroxidation/genetics , Male , Mice , Mice, Inbred C57BL , Obesity/genetics
13.
J Clin Endocrinol Metab ; 96(7): 2089-97, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21543428

ABSTRACT

CONTEXT: GH secretion peaks at puberty and continues to be secreted in adulthood, albeit at a declining rate. Profound GH deficiency (GHD) in adults with pituitary disease is associated with symptoms that improve with GH substitution, but it is important to tailor the GH dose to avoid overtreatment. Measurement of serum IGF-I levels is an important clinical tool in this regard, but it is well recognized that some patients receiving GH treatment do not show an increase in IGF-I. OBJECTIVE: The objective of the study was to identify novel serum biomarkers of GH treatment in adults with GHD. DESIGN AND PATIENTS: Eight patients with profound GHD as a consequence of a pituitary adenoma or its treatment were evaluated before and 3 months after GH replacement therapy (0.2-0.4 mg/d). MAIN OUTCOME MEASURES: Serum proteomic changes were studied using two-dimensional gel electrophoresis and mass spectrometry. Protein profiles were analyzed and compared in serum samples obtained before and after GH treatment. RESULTS: The levels of six serum protein spots were significantly altered after GH substitution. These proteins were identified as five isoforms of haptoglobin (decreased in posttreatment samples) and one isoform of apolipoprotein A-I (increased in posttreatment samples). Importantly, changes in the levels of the identified proteins were associated with decreases in fat mass and increases in lean mass in all patients. These results were independent of serum IGF-I levels. CONCLUSIONS: Evaluation of the identified proteins provides a novel alternative to traditional markers of GH status, such as serum IGF-I levels, to assess GH therapy in GH deficient adults.


Subject(s)
Adenoma/complications , Biomarkers/blood , Hormone Replacement Therapy , Human Growth Hormone/therapeutic use , Hypopituitarism/therapy , Pituitary Neoplasms/complications , Adenoma/blood , Adult , Human Growth Hormone/deficiency , Humans , Hypopituitarism/blood , Hypopituitarism/etiology , Mass Spectrometry , Pituitary Neoplasms/blood , Treatment Outcome
14.
Growth Horm IGF Res ; 21(3): 113-23, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21470887

ABSTRACT

The last two decades have seen resurgence in research focused on adipose tissue. In part, the enhanced interest stems from an alarming increase in obesity rates worldwide. However, an understanding that this once simple tissue is significantly more intricate and interactive than previously realized has fostered additional attention. While few would argue that growth hormone (GH) radically alters fat mass, newer findings revealing the complexity of adipose tissue requires that GH's influence on this tissue be reexamined. Therefore, the objective of this review is to describe the more recent understanding of adipose tissue and to summarize our current knowledge of how GH may influence and contribute to these newer complexities of this tissue with special focus on the available data from mice with altered GH action.


Subject(s)
Adipocytes/metabolism , Adipose Tissue/metabolism , Growth Hormone/metabolism , Animals , Body Composition , Extracellular Matrix , Humans , Mice
15.
J Appl Physiol (1985) ; 110(1): 149-56, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20966191

ABSTRACT

Erythropoietin (Epo) is produced primarily in the kidneys upon low blood oxygen availability and stimulates erythropoiesis in the bone marrow. Recombinant human Epo (rHuEpo), a drug developed to increase arterial oxygen content in patients, is also illicitly used by athletes to improve their endurance performance. Therefore, a robust and sensitive test to detect its abuse is needed. The aim of the present study was to investigate potential human serum biomarkers of Epo abuse employing a proteomic approach. Eight healthy male subjects were injected subcutaneously with rHuEpo (5,000 IU) every second day for a 16-day period. Serum was collected before starting the treatment regime and again at days 8 and 16 during the treatment period. Samples were homogenized and proteins separated by two-dimensional gel electrophoresis (2DE). Spots that changed significantly in response to rHuEpo treatment were identified by mass spectrometry. Both the number of reticulocytes and erythrocytes increased throughout the study, leading to a significant increase in hematocrit and hemoglobin content. In addition, transferrin levels increased but the percentage of iron bound to transferrin and ferritin levels decreased. Out of 97 serum proteins, seven were found to decrease significantly at day 16 compared with pre-Epo administration, and were identified as four isoforms of haptoglobin, two isoforms of transferrin, and a mixture of hemopexin and albumin. In support, total serum haptoglobin levels were found to be significantly decreased at both days 8 and 16. Thus a 2DE proteomic approach for discovery of novel markers of Epo action appears feasible.


Subject(s)
Blood Proteins/analysis , Doping in Sports/prevention & control , Erythropoietin/blood , Performance-Enhancing Substances/blood , Proteome/analysis , Substance Abuse Detection/methods , Adult , Humans , Male , Reproducibility of Results , Sensitivity and Specificity
16.
Eur J Endocrinol ; 164(2): 157-67, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21059862

ABSTRACT

CONTEXT: Transsphenoidal adenomectomy is the primary treatment for acromegaly. However, assessment of the therapeutical outcome remains problematic since the existing biomarkers of disease activity frequently show discordant results. OBJECTIVE: To discover novel serum biomarkers of disease activity in acromegalic patients before and after surgery. DESIGN: Serum samples of eight newly diagnosed acromegaly patients before and after transsphenoidal surgery were analyzed for proteomic changes by two-dimensional gel electrophoresis. Protein spots displaying statistically significant changes, pre- versus post-surgery, were identified by mass spectrometry (MS), tandem MS (MS/MS), and western blot analysis. RESULTS: Six protein spots displaying decreased intensities after surgery were identified as transthyretin (two isoforms), haptoglobin α2, ß-hemoglobin, and apolipoprotein A-1 (two isoforms). One protein spot, identified as complement C4B precursor, was increased after the surgery. CONCLUSIONS: Seven serum protein spots were differentially expressed following surgery in acromegalic patients. The identified proteins represent potential novel biomarkers to assess the effectiveness of surgical treatment in acromegalic individuals. Future studies will validate the use of the identified proteins as biomarkers of disease activity after medical treatment of acromegaly.


Subject(s)
Acromegaly/blood , Adenoma/blood , Pituitary Neoplasms/blood , Proteome/analysis , Acromegaly/surgery , Adenoma/surgery , Adult , Aged , Biomarkers/blood , Blotting, Western , Databases, Factual , Electrophoresis, Gel, Two-Dimensional , Female , Haptoglobins , Human Growth Hormone/blood , Humans , Insulin-Like Growth Factor I , Male , Middle Aged , Pituitary Neoplasms/surgery , Prealbumin , Proteomics/methods , Tandem Mass Spectrometry , Treatment Outcome
17.
Endocr Rev ; 32(3): 356-86, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21123740

ABSTRACT

Disruption of the GH receptor (GHR) gene eliminates GH-induced intracellular signaling and, thus, its biological actions. Therefore, the GHR gene disrupted mouse (GHR-/-) has been and is a valuable tool for helping to define various parameters of GH physiology. Since its creation in 1995, this mouse strain has been used by our laboratory and others for numerous studies ranging from growth to aging. Some of the most notable discoveries are their extreme insulin sensitivity in the presence of obesity. Also, the animals have an extended lifespan, which has generated a large number of investigations into the roles of GH and IGF-I in the aging process. This review summarizes the many results derived from the GHR-/- mice. We have attempted to present the findings in the context of current knowledge regarding GH action and, where applicable, to discuss how these mice compare to GH insensitivity syndrome in humans.


Subject(s)
Receptors, Somatotropin/physiology , Age Factors , Animals , Female , Growth Hormone/physiology , Humans , Insulin Resistance , Insulin-Like Growth Factor I/physiology , Male , Mice , Mice, Knockout , Obesity/physiopathology , Phenotype , Receptors, Somatotropin/deficiency , Receptors, Somatotropin/genetics , Signal Transduction
18.
Growth Horm IGF Res ; 19(6): 471-7, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19386527

ABSTRACT

OBJECTIVE: To identify biomarkers of growth hormone (GH) and insulin-like growth factor 1 (IGF-1) action in human serum. BACKGROUND: The search for new markers of GH activity has received extensive attention given that the current biomarkers (IGF-1, IGFBP-3 and collagen peptides) show substantial variability in the population, and are not reliably predictive of either the physiologic effects of GH therapy or the detection of GH abuse by athletes. GH releasing hormone (GHRH) is a polypeptide synthesized in the hypothalamus that binds to receptors on pituitary somatotropes to promote the synthesis and release of GH. Serum GH and IGF-1 levels have been shown to increase with administration of GHRH or CJC-1295, a long-acting GHRH analog. DESIGN: Sera from 11 healthy young adult men before and one week after CJC-1295 injection were analyzed by two-dimensional gel electrophoresis for proteomic changes. Serum proteins displaying significant changes before and after treatment were subsequently identified using mass spectrometry. In addition, correlations between these proteins and GH or IGF-1 levels were evaluated. RESULTS: Two protein spots that displayed decreased intensities after treatment were identified as an apolipoprotein A1 isoform and a transthyretin isoform. Three protein spots upregulated by CJC-1295 treatment included beta-hemoglobin, a C-terminal fragment of albumin, and a mix of an immunoglobulin fragment and another C-terminal albumin fragment. A linear relationship was found between the spot containing immunoglobulin and albumin fragments and IGF-1 levels. CONCLUSIONS: Although the molecular mechanisms linking the identified proteins to GH and IGF-1 biological activity remain to be clarified, the results suggest that they represent potential biomarkers of GH and/or IGF-1 action.


Subject(s)
Growth Hormone-Releasing Hormone/analogs & derivatives , Growth Hormone/metabolism , Insulin-Like Growth Factor I/metabolism , Peptide Fragments/pharmacology , Adult , Biomarkers/metabolism , Blood Proteins/metabolism , Electrophoresis, Gel, Two-Dimensional/methods , Growth Hormone-Releasing Hormone/pharmacology , Humans , Hypothalamus/metabolism , Male , Mass Spectrometry/methods , Models, Biological , Pituitary Gland/metabolism , Proteomics/methods , Recombinant Proteins/chemistry
19.
Nat Clin Pract Endocrinol Metab ; 3(4): 355-68, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17377618

ABSTRACT

Molecular techniques have had and are continuing to have a strong effect on clinical research and on diagnosis and screening of many endocrine disorders. To undertake research and interpret the results of others, it is important to know how and when to use molecular techniques such as Southern, northern and western blotting and the polymerase chain reaction. Knowledge of the human genome and how genes translate into proteins is required for a full understanding of the burgeoning fields of genomics and proteomics. Genetic manipulation of experimental species, which uses transgenic and gene-knockout technology, has led to important advances in determining the relationship between genes and their encoded proteins' function in the intact organism. This article describes these aspects of molecular biology, and gives specific examples of how they can be applied to clinical endocrinology and metabolism.


Subject(s)
DNA Primers , Endocrinology/methods , Molecular Biology/methods , Animals , Animals, Genetically Modified , Cloning, Molecular , DNA/analysis , DNA Mutational Analysis/methods , Humans , Protein Biosynthesis , Protein Processing, Post-Translational , RNA/analysis , RNA Processing, Post-Transcriptional
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