Insulin-like Growth Factor II Research Articles

Long-term Vaccination with Multiple Peptides Derived from Cancer-Testis Antigens Can Maintain a Specific T-cell Response and Achieve Disease Stability in Advanced Biliary Tract Cancer

The prognosis of patients with advanced biliary tract cancer (BTC) is extremely poor and there are only a few standard treatments. We conducted a phase I trial to investigate the safety, immune response, and antitumor effect of vaccination with four peptides derived from cancer-testis antigens, with a focus on their fluctuations during long-term vaccination until the disease had progressed. Nine patients with advanced BTC who had unresectable tumors and were refractory to standard chemotherapy were enrolled. HLA-A*2402-restricted epitope peptides, lymphocyte antigen 6 complex locus K, TTK protein kinase, insulin-like growth factor-II mRNA-binding protein 3, and DEP domain containing 1 were vaccinated subcutaneously once a week at doses of 0.5, 1, or 2 mg and continued until disease progression. The adverse events were assessed by Common Terminology Criteria for Adverse Events and the immune response was monitored by an enzyme-linked immunospot assay or by flow cytometry. The clinical effects observed were tumor response, progression-free survival (PFS), and overall survival (OS). Four-peptide vaccination was well tolerated. No grade 3 or 4 adverse events were observed. Peptide-specific T-cell immune responses were observed in seven of nine patients and clinical responses were observed in six of nine patients. The median PFS and OS were 156 and 380 days. The injection site reaction and CTL induction seemed to be prognostic factors of both PFS and OS. Four-peptide vaccination was well tolerated and seemed to provide some clinical benefit to some patients. These immunologic and clinical responses were maintained over the long term through continuous vaccinations.

Microstructural Brain and Multivoxel Spectroscopy in Very Low Birth Weight Infants Related to Insulin-Like Growth Factor Concentration and Early Growth

Background: Very low birth weight (VLBW) children have higher risk of neurologic disabilities and growth factors are essential for brain maturation. Aim: To assess whether there are differences in neurologic findings, psychometric parameters and microstructural brain morphology in 1-year-old VLBW infants versus term healthy controls and whether these differences are related to hormonal/growth changes. Methods: Prospective anthropometry, prefeed venous blood sample [insulin, insulin-like growth factor-I (IGF-I), insulin-like growth factor-II (IGF-II), leptin, glucose], neurologic and imaging assessment, at age 1 year in 34 VLBW infants (12 SGA; 10 M) and 10 healthy term controls (5 M). Results: IGF-I concentrations at 1 month of corrected age were 20% lower in SGA versus appropriate for gestational age VLBW (p < 0.02). Gray and white matter volume and fractional anisotropy in 15/27 regions were decreased (p < 0.001). Abnormal spectroscopy was observed in 4 zones in VLBW versus term controls (p < 0.001). Some of these changes were associated with different periods of first-year growth and IGF-I/IGF-II, leptin and HOMA-IR. Conclusions: VLBW infants show differences in brain volumes and microstructural brain morphology as compared to term controls, changes related to circulating growth factor and anthropometry changes in the first year. This apparent reorganization of the developing brain offers a unique opportunity to investigate the relationship between changes in cortical anatomy, cognitive and social impairments and periods of early growth.

Variation in sequences and mRNA expression levels of growth hormone (GH), insulin-like growth factor I (IGF-I) and II (IGF-II) genes between prolific Lezhi black goat and non-prolific Tibetan goat (Capra hircus)

Growth hormone (GH), insulin-like growth factor-I (IGF-I), and II (IGF-II) play a key role in the development of preantral to preovulatory follicles in some species. To better understand the role of these genes in controlling follicular development and fecundity in goats, in the present study, we first cloned the cDNA encoding GH, IGF-I and IGF-II from prolific Lezhi black goat and non-prolific Tibetan goat (Capra hircus), and their mRNA expression between the two breeds were compared. By reverse transcriptase-polymerase chain reaction (RT-PCR) strategy, we obtained full-length 688-bp GH, 493-bp IGF-I, and 566-bp IGF-II cDNAs, encoding for 217 amino acid (aa) GH, 154aa IGF-I, and 179aa IGF-II putative proteins. Analysis of their nucleotide and amino acid sequences revealed a high degree of identity between the two breeds, although one base change in GH resulted in one amino acid substitution in the translated proteins. However, two base changes in IGF-I and IGF-II did not lead to any amino acid changes. Real-time PCR analyses revealed that in the middle of estrus, GH, IGF-I and IGF-II genes were expressed, albeit at different levels, in all three tissues (anterior pituitary, endometrium and ovary) examined. GH was most highly expressed in ovary (P<0.01) and its expression was greater in all three tissues examined in Lezhi black goat than in Tibetan goat (P<0.05). IGF-I and IGF-II genes were expressed at a higher (P<0.05) level in anterior pituitary of Lezhi black goat than that in Tibetan goat, but they had a similar expression pattern in endometrium and ovary. These results provide the foundation of information required for future studies of these gene effects on goat fecundity.

Regulation of integrin αV subunit expression by sulfatide in hepatocellular carcinoma cells

Integrin is important in migration and metastasis of tumor cells. Changes of integrin expression and distribution will cause an alteration of cellular adhesion and migration behaviors. In this study, we investigated sulfatide regulation of the integrin αV subunit expression in hepatoma cells and observed that either exogenous or endogenous sulfatide elicited a robust upregulation of integrin αV subunit mRNA and protein expression in hepatoma cells. This regulatory effect occurred with a corresponding phosphorylation (T739) of the transcription factor Sp1. Based on the electrophoretic mobility shift assay, sulfatide enhanced the integrin αV promoter activity and strengthened the Sp1 complex super-shift. The results of chromatin immunoprecipitation analysis also indicated that sulfatide enhanced Sp1 binding to the integrin αV promoter in vivo. Silence of Sp1 diminished the stimulation of integrin αV expression by sulfatide. In the early stage of sulfatide stimulation, phosphorylation of Erk as well as c-Src was noted, and inhibition of Erk activation with either U0126 or PD98059 significantly suppressed Sp1 phosphorylation and integrin αV expression. We demonstrated that sulfatide regulated integrin αV expression and cell adhesion, which was associated with Erk activation.

Abnormal expression of insulin-like growth factor-II and intervening of its mRNA transcription in the promotion of HepG2 cell apoptosis

To explore the expression and pathological features of insulin-like growth factor-II (IGF-II) in tissues and sera of hepatocellular carcinoma (HCC) patients and the siRNA-mediated inhibition of IGF-II mRNA transcription in human HepG2 cells. From December 2009 to August 2010, the self-control HCC, paracancerous and distal cancerous tissues were collected to analyze the expression of IGF-II. The serum levels of IGF-II expression were detected for pathological features. IGF-II expression in HepG2 cells was intervened by siRNA. IGF-II mRNA or IGF-II level and analyzed by reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR or enzyme-linked immunosorbent assay (ELISA). And the ratio of cell apoptosis was analyzed by EdU/Hoechst33342. The levels of IGF-II expression in HCC tissues at mRNA (100%, 30/30) or protein (83.3%, 25/30) were significantly higher (P < 0.01) than those in para-cancerous (46.7%, 53.3%) or distal cancerous tissues (0, 0). The serum level of IGF-II was significantly higher in HCC patients (3.74 ± 0.67) ng/L than that in cases with benign liver diseases (1.93 ± 0.17) ng/L and controls (1.14 ± 0.14) ng/L (P < 0.001). The expression of IGF-II in the HCC group was associated with HBV infection (t = 5.390, P < 0.001). After siRNA transfection, the expression of IGF-II decreased significantly in HepG2 cells at mRNA or protein levels. The down-regulated expression of IGF-II was dependent on the dose and time of IGF-II siRNA. And the apoptotic index of HepG2 cells and the sensitivity to adriamycin both increased. The expression of IGF-II is closely associated with the progression of HCC. And the intervening of its transcription may promote apoptosis and sensitize to adriamycin.

Insulin-like growth factor-II is produced by, signals to and is an important survival factor for the mature podocyte in man and mouse.

Podocytes are crucial for preventing the passage of albumin into the urine and, when lost, are associated with the development of albuminuria, renal failure and cardiovascular disease. Podocytes have limited capacity to regenerate, therefore pro-survival mechanisms are critically important. Insulin-like growth factor-II (IGF-II) is a potent survival and growth factor; however, its major function is thought to be in prenatal development, when circulating levels are high. IGF-II has only previously been reported to continue to be expressed in discrete regions of the brain into adulthood in rodents, with systemic levels being undetectable. Using conditionally immortalized human and ex vivo adult mouse cells of the glomerulus, we demonstrated the podocyte to be the major glomerular source and target of IGF-II; it signals to this cell via the IGF-I receptor via the PI3 kinase and MAPK pathways. Functionally, a reduction in IGF signalling causes podocyte cell death in vitro and glomerular disease in vivo in an aged IGF-II transgenic mouse that produces approximately 60% of IGF-II due to a lack of the P2 promoter of this gene. Collectively, this work reveals the fundamental importance of IGF-II in the mature podocyte for glomerular health across mammalian species.

Insulin-Like Growth Factor-II (IGF-II) Prevents Proinflammatory Cytokine-Induced Apoptosis and Significantly Improves Islet Survival After Transplantation

The early loss of functional islet mass (50-70%) due to apoptosis after clinical transplantation contributes to islet allograft failure. Insulin-like growth factor (IGF)-II is an antiapoptotic protein that is highly expressed in β-cells during development but rapidly decreases in postnatal life. We used an adenoviral (Ad) vector to overexpress IGF-II in isolated rat islets and investigated its antiapoptotic action against exogenous cytokines interleukin-1β- and interferon-γ-induced islet cell death in vitro. Using an immunocompromised marginal mass islet transplant model, the ability of Ad-IGF-II-transduced rat islets to restore euglycemia in nonobese diabetic/severe combined immunodeficient diabetic recipients was assessed. Ad-IGF-II transduction did not affect islet viability or function. Ad-IGF-II cytokine-treated islets exhibited decreased cell death (40% ± 2.8%) versus Ad-GFP and untransduced control islets (63.2% ± 2.5% and 53.6% ± 2.3%, respectively). Ad-IGF-II overexpression during cytokine treatment resulted in a marked reduction in terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive apoptotic cells (8.3% ± 1.4%) versus Ad-GFP control (41% ± 4.2%) and untransduced control islets (46.5% ± 6.2%). Western blot analysis confirmed that IGF-II inhibits apoptosis via activation of the phosphatidylinositol 3-kinase/Akt signaling pathway. Transplantation of IGF-II overexpressing islets under the kidney capsule of diabetic mice restored euglycemia in 77.8% of recipients compared with 18.2% and 47.5% of Ad-GFP and untransduced control islet recipients, respectively (P<0.05, log-rank [Mantel-Cox] test). Antiapoptotic IGF-II decreases apoptosis in vitro and significantly improved islet transplant outcomes in vivo. Antiapoptotic gene transfer is a potentially powerful tool to improve islet survival after transplantation.

Expression of the oncofetal protein IGF2BP3 in endometrial clear cell carcinoma: assessment of frequency and significance

Insulin-like growth factor-II messenger RNA-binding protein 3 (IGF2BP3 or IMP3) is a biomarker whose expression has been found to be a negative prognostic factor in several neoplasms including ovarian clear cell carcinoma (CCC). In this study, we analyzed the frequency and clinicopathologic significance of IMP3 expression, as assessed by immunohistochemistry and as scored using a modified H-score system, in a cohort of 50 endometrial CCCs. Cases with scores of 0 to 100, 101 to 200, and 201 to 300 were classified as negative/mildly positive (n = 17), moderately positive (n = 20), and strongly positive (n = 13), respectively. A distinctive pattern of increased staining at the myoinvasive front (relative to the main tumor) was evident in 46% of the cases with evaluable foci of myometrial invasion. Moderate/strong IMP3 staining was associated with a tumor architectural pattern that has been reported to be of poor prognostic significance: at least 10% of the tumor composed of solid architecture or individual infiltrating tumor cells (P = .01). Increasing levels of IMP3 expression showed a trend toward decreasing relapse-free survival (RFS; median survival, 75.6, 81.3, and 48.4 months for the negative/mildly, moderately, and strongly positive groups, respectively [P = .09]). However, IMP3 expression was not significantly associated with reduced overall survival or RFS in a multivariate analytic model. The finding in a subset of our cases of increased IMP3 expression at the tumoral myoinvasive front is consistent with a role for IMP3 in invasiveness, as is the trend toward reduced RFS in cases expressing IMP3 at high levels. These preliminary findings suggest that IMP3 expression may be involved in the pathogenesis of CCC and is worthy of further exploration.

Insulin-like growth factor II (IGF-II) and follicle stimulating hormone (FSH) combinations can improve the in vitro development of grown oocytes enclosed in caprine preantral follicles

ObjectiveEvaluate the possible role of IGF-II alone or in association with FSH on in vitro development of isolated caprine preantral follicles. MethodsPreantral follicles (≥150μm) were isolated from goat ovaries and cultured for 18days in basic αMEM medium (control) or supplemented with IGF-II alone at 20 or 50ng/ml, named IGF20 and IGF50, respectively, or in combination with recombinant FSH (FSH, IGF20F or IGF50F). During in vitro culture, the follicles were analyzed by using morphology criteria, antrum formation and growth rate as parameters. After 18days of follicular culture, oocytes equal to or larger than 110μm were used for in vitro maturation (IVM). Oocyte viability and meiosis resumption were assessed by fluorescence microscopy after labeling with calcein-AM, ethidium homodimer and Hoechst 33342. ResultsThe IGF20 treatment was the only treatment capable of maintaining the percentage of morphologically normal follicles from D0 until D6 and from D12 to D18 (p>0.05), while in all other treatments the percentage of morphologically normal follicles decreased progressively during 18days of in vitro culture (p<0.05). At D18, all treatments with IGF-II or FSH resulted in a significantly higher percentage of normal follicles when compared to αMEM alone. The IGF50F treatment provided a significantly higher early antrum formation rate when compared to αMEM and FSH alone. The addition of IGF-II alone (20 or 50ng/ml) or in combination with FSH prevented oocyte degeneration after IVM. Moreover, the FSH treatment demonstrated a lower percentage of oocyte degeneration when compared to control (4.35% vs. 26.3%, respectively; p<0.05). Regarding meiosis resumption, the IGF20F treatment was the only treatment that significantly differed from αMEM alone. All treatments except the control (αMEM alone) presented oocytes at metaphase II. ConclusionIGF-II associated with FSH stimulated in vitro follicular development, oocyte viability and meiotic resumption of caprine oocytes after IVM.

Biochemical Characterization of Individual Human Glycosylated pro-Insulin-like Growth Factor (IGF)-II and big-IGF-II Isoforms Associated with Cancer

Insulin-like growth factor II (IGF-II) is a major embryonic growth factor belonging to the insulin-like growth factor family, which includes insulin and IGF-I. Its expression in humans is tightly controlled by maternal imprinting, a genetic restraint that is lost in many cancers, resulting in up-regulation of both mature IGF-II mRNA and protein expression. Additionally, increased expression of several longer isoforms of IGF-II, termed "pro" and "big" IGF-II, has been observed. To date, it is ambiguous as to what role these IGF-II isoforms have in initiating and sustaining tumorigenesis and whether they are bioavailable. We have expressed each individual IGF-II isoform in their proper O-glycosylated format and established that all bind to the IGF-I receptor and both insulin receptors A and B, resulting in their activation and subsequent stimulation of fibroblast proliferation. We also confirmed that all isoforms are able to be sequestered into binary complexes with several IGF-binding proteins (IGFBP-2, IGFBP-3, and IGFBP-5). In contrast to this, ternary complex formation with IGFBP-3 or IGFBP-5 and the auxillary protein, acid labile subunit, was severely diminished. Furthermore, big-IGF-II isoforms bound much more weakly to purified ectodomain of the natural IGF-II scavenging receptor, IGF-IIR. IGF-II isoforms thus possess unique biological properties that may enable them to escape normal sequestration avenues and remain bioavailable in vivo to sustain oncogenic signaling.

Genetic polymorphisms of the IGF-II gene intron 8 coding region and its association with growth and carcass traits in yak

Insulin-like growth factor II (IGF-II) plays a key role in mammalian growth and is involved in stimulating fetal cell division, differentiation, and metabolic regulation. IGF-II is considered a candidate gene for genetic markers of growth and carcass traits. Therefore, in this study, the associations of single nucleotide polymorphisms (SNPs) in the IGF-II gene region with growth and carcass characteristics in five yak breeds were investigated. Two SNPs, G(330)C and A(358)G, were identified by sequencing intron 8 of the IGF-II gene in homozygotes. Two alleles, A and B, and three genotypes, AA, AB, and BB, were identified by polymerase chain reaction. Genotypic frequencies of IGF-II allele B were 0.8623, 0.8936, 0.8535, 0.8676, and 0.8300 for Datong yak, Gannan yak, Tianzhu white yak, Qinghai Plateau yak, and Xinjiang yak, respectively. Allele and the genotype of IGF-II were strongly associated with growth and carcass traits. Least square analysis revealed a significant effect (P < 0.01) of genotypes AA and AB compared with genotype BB on live-weight (at 12, 13-24, and 25-36 months of age), average daily weight gain (P < 0.01) and carcass weight (P < 0.05). Animals with genotype AB had a higher mean rib eye area, and a lower mean yield grade. The results indicated that the IGF-II gene acts by a primarily additive biological mechanism by adding weight independently of skeletal growth.

Gastric Neuroendocrine Carcinoma with Non-islet Cell Tumor Hypoglycemia Associated with Enhanced Production of Insulin-like Growth Factor II

A 75-year-old man was admitted to the hospital with a loss of consciousness. His blood glucose level was 24 mg/dL. Abdominal computed tomography revealed multiple metastatic lesions in the liver, while upper endoscopy disclosed advanced gastric cancer. The hypoglycemia was refractory despite the administration of glucose and steroid therapy. The patient died within one month of admission. An autopsy revealed neuroendocrine-type gastric cancer, which, on examination with immunohistochemistry, was found to be negative for insulin and insulin-like growth factor I and positive for insulin-like growth factor II (IGF-II). The patient was diagnosed as having gastric cancer with non-islet cell tumor hypoglycemia (NICTH) caused by IGF-II.

Epimedium Flavonoids Counteract the Side Effects of Glucocorticoids on Hypothalamic-Pituitary-Adrenal Axis

Our previous studies demonstrated that the epimedium herb, when simultaneously used with GCs, counteracted suppressive effects of GCs on the HPA axis without adverse influence on the therapeutic action of GCs. Here, total flavones were extracted from the epimedium flavonoids (EFs) and then used to investigate whether EFs provide protective effects on the HPA axis. We found that GCs induced a significant decrease in body weight gain, adrenal gland weight gain, and plasma adrenocorticotropin (ACTH) and corticosterone levels. After treatment with EFs, body weight gain, adrenal gland weight gain, and plasma corticosterone level were significantly restored, whilst plasma ACTH level was partially elevated. EFs were also shown to promote cell proliferation in the outer layer of adrenal cortex and to enhance the migration of newly divided cells toward the inner layer. To elucidate the underlying mechanisms, the mRNA expression of insulin-like growth factor II (IGF-II) was measured, and EFs significantly upregulated IGF-II expression. Our results indicated that EFs counteract the suppression of the HPA axis induced by GCs. This may involve both the ACTH and IGF-II pathways and thereby promote regeneration of the adrenal cortex suggesting a potential clinical application of EFs against the suppressive effects of GCs on the HPA axis.

Increased Intragenic IGF2 Methylation is Associated with Repression of Insulator Activity and Elevated Expression in Serous Ovarian Carcinoma

Overexpression of insulin-like growth factor-II (IGF2) is a prominent characteristic of many epithelial ovarian malignancies. IGF2 imprinting and transcription are regulated in part through DNA methylation, which in turn regulates binding of the insulator protein CTCF within the IGF2/H19 imprint center. We have shown that IGF2 overexpression in ovarian cancer is associated with hypermethylation of CTCF binding sites within the IGF2/H19 imprint center. The aim of this study was to investigate the methylation and binding capacity of a novel putative CTCF binding motif located intragenic to IGF2 and determine how this relates to IGF2 expression. Among 35 primary serous epithelial ovarian cancer specimens, methylation of two CpGs, including one within the core binding motif and another adjacent to this motif, was higher in the 18 cancers with elevated IGF2 expression versus 10 with low expression (average 68.2 versus 38.5%; p < 0.0001). We also found that the CpG site within the CTCF binding motif is hypermethylated in male gametes (>92%; average 93.2%; N = 16). We confirmed binding of CTCF to this region in ovarian cancer cells, as well as the paralog of CTCF, Brother Of the Regulator of Imprinted Sites (BORIS), which is frequently overexpressed in cancers. The unmethylated CTCF binding motif has insulator activity in cells that express CTCF or BORIS, but not in cells that express both CTCF and BORIS. These intragenic CpG dinucleotides therefore comprise a novel paternal germline imprint mark and are located in a binding motif for the insulator protein CTCF. Methylation of the CpG dinucleotides is positively correlated with IGF2 transcription, indicating that increased methylation represses insulator function. These combined results suggest that methylation and CTCF binding at this region play important roles in regulating the level of IGF2 transcription. Our data have revealed a novel epigenetic regulatory element within the IGF2/H19 imprinted domain that is highly relevant to aberrant IGF2 expression in ovarian malignancies.

Remarkable body weight gain in a patient in the terminal phase of gastric carcinoma secondary to ‘big IGF-II’— producing tumor

Weight loss as a result of cachexia with malignancy is frequently encountered in clinical practice. However, previous reports have not demonstrated weight gain in terminal phase of malignancy. We report a 69-year-old male admitted with an exacerbation of interstitial pneumonia and multiple metastases of relapsed gastric adenocarcinoma complicated with refractory hypoglycemia. He had gained approximately 10 kg weight with an acromegaloid appearance in a one-month period before admission. The presence of big insulin-like growth fact II (Big IGF-II) in his blood and an immunohistochemical study on the tumor cells confirmed a diagnosis of non-islet cell tumor hypoglycemia. This report possibly illustrates an unreported differential diagnosis of weight gain in terminal phase of malignancy.

The roles of Insulin-like growth factor-II messenger RNA (mRNA)-binding protein-3 (IMP-3) in melanoma progression

Insulin-like growth factor-II messenger RNA (mRNA)-binding protein-3 (IMP-3) is a member of insulin-like growth factor-II mRNA-binding protein family and is expressed during embryogenesis and in some malignancies. Recently, IMP-3 has been found to be a novel progression marker in malignant melanoma.

Chemical Synthesis of a Fluorescent IGF-II Analogue

Insulin-like growth factor II (IGF-II) is a protein with high structural and sequence similarity to insulin. Unlike insulin, it binds both the type 1 IGF receptor and the exon 11- isoform of the insulin receptor with high affinity. The overexpression and up regulation of IGF-II has been associated with the progression of various forms of cancer. The exact binding mechanism of IGF-II to its high affinity receptors is still not completely understood. Herein we describe the successful synthesis of a novel fluorescent IGF-II protein (F19Cou IGF-II), where residue 19 (phenylalanine) has been replaced by a fluorescent chromophore (coumaryl glycine). This novel coumaryl IGF-II analogue will be a useful tool for analysing the receptor interaction mechanisms in future studies.

A case report of non-islet cell tumour hypoglycaemia associated with ovarian germ-cell tumour

Background: Non-islet cell tumour hypoglycaemia is a rare paraneoplastic condition in which tumours secrete a high-molecular-weight precursor of insulin-like growth factor-II causing hypoglycaemia and can be difficult to identify and treat. Case Presentation: This is the case of a 27-year-old patient from Africa with metastatic ovarian yolk sac tumour who presented with hypoglycaemia and was subsequently diagnosed with non-islet cell tumour hypoglycaemia. Case Management: Our patient required higher doses of glucocorticosteroids than reported in the literature in combination with recombinant growth hormone therapy in order to control her hypoglycaemia. Case Outcome: This is the first case of non-islet cell tumour hypoglycaemia described in association with a germ-cell tumour. Her management required collaboration between the endocrinology team, the palliative care team, the acute medicine team and physicians in Africa to enable her safe journey home. Conclusions: This case illustrates the need for awareness among general physicians of rare tumour manifestations and the need for multidisciplinary input for the optimal management of these patients.

Age-related changes in serum levels of insulin-like growth factor-II and its binding proteins correlate with calcaneal bone mineral density among post-menopausal South-Indian women

BackgroundInsulin-like growth factor (IGF) system components are important regulators of bone metabolism, which have a predominant role in determining bone mineral density (BMD). While the serum levels of IGF-I are regulated by various systemic hormones and growth factors, IGF-II levels reflect the skeletal production relative to physical activity, mechanical loading, aging, race etc. Though various studies have been carried out among women of different ethnic groups to understand the relationship between serum levels of IGF-II and BMD, the results seem to be quite inconclusive. MethodsWe evaluated the same, recruiting South-Indian women who engage themselves in a wide variety of physical activities pertaining to their profession and life style. ResultsSerum levels of IGF-II and IGF binding protein (IGFBP)-3 showed positive correlation with calcaneal BMD, whereas IGFBP-4 showed negative correlation. These IGF system components exhibited similar correlations with serum bone formation markers and opposite trend with bone resorption marker. While both IGF-II and IGFBP-3 levels were observed to be decreased with aging and menopause, IGFBP-4 levels increased. ConclusionsThe alterations in serum levels of IGF-II and its binding proteins due to aging and menopause could be some of the major contributors of decreased calcaneal BMD observed among elderly women.

O1-077 - Cancer Vaccination Trial with Novel Multiple Peptides in Previously Treated Advanced Non-Small-Cell Lung Cancer

ABSTRACT Background Cell division cycle associated gene 1 (CDCA1) and insulin-like growth factor-II mRNA binding protein 3 (IMP-3) are novel cancer-testis antigen overexpressed in non-small-cell lung cancer (NSCLC). The aim of this study was to evaluate the safety, efficacy and immunological response of a novel peptide vaccination therapy, CDCA1 and IMP-3, in patients with previously treated advanced NSCLC. Patients and methods Human histocompatibility leukocyte (HLA)-A2402-positive advanced NSCLC patients who failed to standard therapy were enrolled if they had a performance status of three or less, were 80 years or younger and had adequate organ function. The cocktail of two peptides was subcutaneously injected with 1 ml of incomplete Freunds adjuvant to axillary or inguinal regions weekly. Immunological response was evaluated with enzyme-linked immunospot assay before and after vaccinations every 4 weeks. Results From November 2009 through December 2011, 20 patients (13 men and 7 women; median age, 65 years; range, 36–78 years) were enrolled. CDCA1 and IMP-3 specific cytotoxic T lymphocyte responses were observed after vaccinations in 60% and 73% of patients, respectively. The overall response rate was 5% (95% CI, 0.1–24.9%) and stable disease rate was 55% (95% CI, 31.5–76.9%). The median survival time from the start of vaccination therapy was 7 months (range 1–26 months). The median progression-free survival time was 3 months (range 1–15 months). One patient with interstitial lung disease (ILD) had acute exacerbation of ILD during vaccination therapy. The other 19 patients did not have any grade 3 or 4 adverse events. Conclusion This vaccination therapy was effective and well tolerated in patients with NSCLC who failed to standard therapy, and therefore warrants further clinical studies.