Insulin-like Growth Factor Receptor Research Articles

Abstract 3635: Insulin-like growth factor receptor predicts poor prognosis in early-stage non-small cell lung cancer

Abstract Introduction: Insulin like growth factor receptor (IGF1R) is involved in the proliferation and metastasis of various cancers. However, the clinical significance of IGF1R in non-small cell lung cancer (NSCLC) remains to be unclear. In this study, we retrospectively evaluated the correlation between IGF1R expression and clinicopathological features of NSCLC. Method: A total of 782 specimens of NSCLC patients who underwent surgical resection of primary lung cancer between 2010 and 2019 at Osaka Metropolitan University Hospital was enrolled in this study. Immunohistochemical staining of IGF1R were performed, and the correlation between IGF1R expression and clinicopathological features of NSCLC, especially in recurrence free survival (RFS) and overall survival (OS), was evaluated. We also evaluated an association between IGF1R and programmed death ligand 1 (PD-L1) expression. Results: IGF1R expression was positive in 279 (35.8%) patients. IGF1R-positive group had significantly longer smoking histories (median smoking index, 1,000 vs 600, p <0.01) and a higher proportion of squamous cell carcinomas (35.1% vs 14.1 %, p <0.01). IGF1R-positive group also significantly expressed PD-L1 (24.4% vs 6.4%, p<0.01). IGF1R-positive group had significantly worse RFS (10-year recurrence-free rate; 42.7% vs 53.7%, p <0.01) and OS (10-year survival rate; 54.4% vs 67.3%, p <0.01). RFS and OS were significantly worse in the IGF1R-positive NSCLC patients at stage 0 and stage I (10-year recurrence-free rate; 49.0% vs 62.9%, p <0.01, 10-year survival rate; 57.4% vs 74.2%, p<0.01), but not in stage II (10-year recurrence-free rate; 52.0% vs 43.8%, p=0.68, 10-year survival rate; 63.4% vs 47.5%, p=0.95) and III (10-year recurrence-free rate; 20.7% vs 20.8%, p=0.77, 10-year survival rate; 38.3% vs 54.6%, p=0.18). IGF1R expression was associated with poorer RFS and OS in early-stage NSCLC. Conclusion: IGF1R is a predictor of poor prognosis in resected early-stage NSCLC. Citation Format: Hiroaki nagamine, Masakazu Yashiro, Megumi Mizutani, Akira Sugimoto, Yoshiya Matsumoto, Yoko Tani, Kenji Sawa, Kazuhiro Yamada, Hiroyasu Kaneda, Tetsuya Watanabe, Kazuhisa Asai, Satoshi Suzuki, Tomoya Kawaguchi. Insulin-like growth factor receptor predicts poor prognosis in early-stage non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3635.

Growth indices and vitamin D content in children with idiopathic short stature depending on the variants of the Taql polymorphism genotype of the VDR gene

Background. The vitamin D receptor (VDR) gene is a key regulator of biological processes, including tissue and cell growth and differentiation, bone metabolism, and modulation of immune responses. Several studies have shown that 1,25(OH)2D increases circulating insulin-like growth factor-1 (IGF-1) levels in vivo and in bone cell cultures. It also increases the expression of IGF-1 receptors in growth plate chondrocytes and several IGF-binding proteins in osteoblasts and bone marrow stromal cell cultures. A vitamin D receptor gene polymorphism is associated with adult height and may affect the growth indicators in children with idiopathic short stature. The purpose was to study the growth indicators and vitamin D levels in children with idiopathic short stature depending on the variants of the Taql polymorphism genotype of the VDR gene. Materials and methods. Determination of the Taql polymorphism of the VDR gene (rs731236) was performed using polymerase chain reaction followed by an analysis of the length of the restriction fragments when detected by agarose gel electrophoresis in 35 children with idiopathic short stature. Results. Most children (68.57 %) were heterozygotes, 17.14 % were homozygotes for T/T alleles, and 14.29 % were homozygotes for C/C alleles. The highest growth retardation has been observed in homozygous T/T carriers. Children carrying homozygotes for C/C alleles had the least growth retardation. Growth hormone levels after clonidine stimulation test and IGF-1 levels in blood were within normal limits in all patients. Vitamin D deficiency was found in T/T homozygotes and vitamin D insufficiency in C/T heterozygotes and C/C homozygotes. Conclusions. Among children with idiopathic short stature, the prevalence of the T/C genotype was 68.57 %. The greatest growth retardation occurred in patients with homozygous T/T alleles (SDS = –2.61 ± 0.31). A decrease in serum vitamin D levels was detected in all children. Vitamin D deficiency (43.83 ± 6.47 nmol/l) was found in children homozygous for the T/T allele, and vitamin D insufficiency — in T/C heterozygotes (58.97 ± 11.78 nmol/l) and C/C homozygotes (56.93 ± 19.54 nmol/l).

Unveiling Modern Strategies for Diagnosing and Treating Grave Orbitopathy

Graves' Orbitopathy (GO), also known as thyroid eye disease or thyroid-associated orbitopathy is characterized by inflammation, ocular muscle hypertrophy, adipogenesis, and oedema (due to glycosaminoglycan accumulation). This condition leads to remodeling, tissue expansion, and/or fibrosis within the fibroadipose tissue or extraocular muscles of the orbit. GO manifests as an extrathyroidal aspect of autoimmune thyroid diseases in both Grave's disease (GD) and Hashimoto's thyroiditis.
 The pathophysiological foundation of GO entails the infiltration of B cells, T cells, and CD34+ fibroblasts in the orbit. B cells generate IGF (insulin-like growth factor), wherein IGF and TRab (Thyroid-Receptor antibodies) stimulate the IGF receptor complex and thyrotropin receptor (respectively) on the surface of CD34+ cells which triggers orbital tissue expansion, orbital protrusion, optic nerve compression, and eyeball displacement, resulting in exophthalmos.
 GO progress through an active phase (characterized by inflammation with visible manifestations), followed by a plateau phase (stabilization of GO manifestations), and a gradual resolution of distinctive residual signs and symptoms (inactive phase). This entire process spans between 18-24 months in untreated patients, where disease manifestations significantly depend on the phase during which the disease is identified.
 GO therapy is intended to shorten the active phase and supress its residual eye manifestations during the inactive phase. In general, GO therapy is categorized into general and disease severity-specific approaches.
 GO therapy often falls short of providing satisfactory results, prompting need of surgery to address lingering clinical manifestations. This review presents the latest insights into the pathogenesis and treatment of GO for better management and outcomes.

Abstract B040: Trispecific killer engagers against IGF1R and fetal form of insulin receptor induce human natural killer cell mediated killing of hormone receptor positive breast cancer in vitro and in vivo

Abstract Patients with hormone receptor-positive (HR+) breast cancers frequently recur with metastatic disease 5-20 years after the initial diagnosis and completion of chemotherapy and prolonged treatment with hormonal therapy. Thus, patients with metastatic HR+ breast cancer need new therapeutic options. A potential mechanism of therapeutic resistance in HR+ patients is the presence of slow-growing, well-differentiated cancer cells that result in clinically detectable metastases after a prolonged latency. One way to eliminate these cancer cells is with immunotherapy (IT). Unfortunately HR+ breast cancers have not responded to current T-cell based IT. The objective of this study is to target HR+ breast cancers using a natural killer (NK) cell based approach that targets cell surface receptors in HR+ breast cancer. The type I IGF receptor (IGF1R) is expressed on HR+ breast cancers and its expression is tightly related to ER function. Endocrine therapy resistance is associated with increased expression of the A-isoform or fetal form of IR (IR-A) and depend on IR signaling. We hypothesized that a drug that would target NK cells to IGF1R or IR-A expressing HR+ cells will be effective in killing them. We generated IGF1R or IR-A specific Tri specific Killer Engagers (TriKE) that leverage expression of IGF1R and IR-A on HR+ cells to cause NK directed death. The IGF1R TriKE consists of humanized single domain antibody (VHH) sequence of a CD16 nanobody to engage and activate NK cells, a humanized VHH of a nanobody against IGF1R to target HR+ breast cancer cells, and an IL-15 molecule between them to drive NK cell expansion and survival. The IGF1R TriKE did not bind embryonal fibroblasts from IGF1R knock out mice (R- cells) but bound R-/IGF1R cells (R- cells engineered to express human IGF1R) and to IGF1R+ MCF-7 and T47D breast cancer cells indicating specific binding to IGF1R on cells. It did not cause downregulation of IGF1R levels in multiple HR+ breast cancer cells and did not inhibit signaling via IGF1R. It specifically induced NK cell degranulation, measured by surface CD107a expression and intracellular IFNγ, against IGF1R+ target breast cancer cells (MCF-7, T-47D, ZR-75-1) in a functional assay compared to the anti-IGF1R nanobody. It did not induce degranulation of NK cells against IGF1R negative cells suggesting that the functionality was specific for IGF1R expressing targets. The IGF1R TriKE enhanced cell killing of HR+ breast cancer cells as measured by cytotoxicity assays. IGF1R TriKE combined with human NK cells inhibited tumor growth of MCF-7 tumors in NSG mice compared to control groups. Human NK cells persisted in peripheral blood three weeks after final NK infusion based on flow cytometry assessment. The IR-A TriKE incorporated a novel engineered non-immunoglobulin IR-A specific peptide binder. Our preliminary data show that the IR-A TriKE degranulated NK cells against breast cancer cells overexpressing IR-A. These data indicate that this strategy could be an effective treatment for HR+ metastatic breast cancer and make them responsive to IT. Citation Format: Courtney Baar, Emily Chiu, Yvette Soignier, Behiye Kodal, Jeffrey S Miller, Martin Felices, Deepali Sachdev. Trispecific killer engagers against IGF1R and fetal form of insulin receptor induce human natural killer cell mediated killing of hormone receptor positive breast cancer in vitro and in vivo [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr B040.

Clinical characteristics of and growth hormone treatment effects on short stature with type 1 insulin-like growth factor receptor (IGF1R) gene alteration.

Short stature with IGF-1 receptor (IGF1R) gene alteration is known as small-for-gestational-age (SGA) short stature with elevated serum IGF1 levels. Its prevalence and clinical characteristics remain unclear. No adapted treatment is available for short stature related to IGF1R gene alteration in Japan, and genetic testing is not yet widely accessible. We investigated short stature with IGF1R gene alterations and analyzed the clinical data of 13 patients using the results of questionnaires issued to the Japanese Society for Pediatric Endocrinology. Four cases were caused by a deletion of chromosome 15q26.3, and eight were caused by heterozygous pathogenic variants in the IGF1R gene. Cases with deletions showed a more severe degree of growth impairment (-4.5 ± 0.43 SD) than those caused by pathological variants (-2.71 ± 0.15 SD) and were accompanied by neurodevelopmental delay. However, cases caused by pathological variants lacked distinctive features. Only three of the 12 cases demonstrated serum IGF1 values exceeding +2 SD, and the other three had values below 0 SD. Four patients did not meet the criteria for SGA at birth. Six patients received GH therapy for SGA short stature and showed improvement in growth rate without any side effects or elevated serum IGF1 levels during treatment. Elevated IGF1 levels (over +2 SD) after GH treatment should be considered a suspicious finding. Owing to the lack of distinctive features, there was a possibility of undiagnosed cases of this condition. Promoting genetic testing and clinical trials on GH administration for this condition is recommended.

Self-Assembling Peptides with Insulin-Like Growth Factor Mimicry.

Growth factor (GF) mimicry involves recapitulating the signaling of larger molecules or cells. Although GF mimicry holds considerable promise in tissue engineering and drug design applications, difficulties in targeting the signaling molecule to the site of delivery and dissociation of mimicking peptides from their target receptors continue to limit its clinical application. To address these challenges, we utilized a self-assembling peptide (SAP) platform to generate synthetic insulin-like growth factor (IGF)-signaling, self-assembling GFs. Our peptide hydrogels are biocompatible and bind target IGF receptors in a dose-dependent fashion, activate proangiogenic signaling, and facilitate formation of angiogenic microtubules in vitro. Furthermore, infiltrated hydrogels are stable for weeks to months. We conclude that the enhanced targeting and long-term stability of our SAP/GF mimicry implants may improve the efficacy and safety of future GF mimic therapeutics.

Correlation analysis between polymorphism of leptin and IGFI genes and body measurements in Barki and Farafra sheep

BackgroundGrowth performance, which can be evaluated using various body measurements, is crucial for providing red meat for human consumption. This study aims to improve the growth performance of sheep by identifying the different genotypes of leptin and IGFI genes associated with higher body measurements.MethodsIn this study, we collected blood samples from Barki (n = 30) and Farafra (n = 30) sheep, and measured their body height, width, length, and weight, and tail width and length to assess their growth performance. We used single strand conformation polymorphism polymerase chain reaction (SSCP-PCR) and sequencing analysis to identify single nucleotide polymorphisms (SNPs) and different genotypes of the leptin and insulin-like growth factor receptor (IGFI) genes associated with sheep body measurements.ResultsThe leptin gene (209 bp) and IGFR gene (345 bp) were found to have three (TT, TG, and GG) and (TT, TC, and CC) genotypes, respectively, by SSCP-PCR. Sequence analysis revealed a SNP in the leptin gene (200G > T) with two alleles (G allele: OR058867 and T allele: OR058868). This SNP resulted in the substitution of the amino acid valine with leucine, which increased the width, length, and weight of the body and tail width in the heterozygous form (TG). The IGFI gene had three SNPs: 305 G > T in TT genotype (OR260670), 184C > T, 265T > C, and 305G > T in TC genotype (OR260669), and 265T > C in CC genotype (OR260671). The TC genotype of the IGFR gene was correlated with higher values of body measurements. Barki sheep were found to have the highest values for body weight, width, length, and tail width compared to Farafra sheep.ConclusionWe recommend the use of entire Barki sheep in inbreeding programs to improve growth performance in Egyptian sheep.

Signaling Pathways Involved in Manganese-Induced Neurotoxicity.

Manganese (Mn) is an essential trace element, but insufficient or excessive bodily amounts can induce neurotoxicity. Mn can directly increase neuronal insulin and activate insulin-like growth factor (IGF) receptors. As an important cofactor, Mn regulates signaling pathways involved in various enzymes. The IGF signaling pathway plays a protective role in the neurotoxicity of Mn, reducing apoptosis in neurons and motor deficits by regulating its downstream protein kinase B (Akt), mitogen-activated protein kinase (MAPK), and mammalian target of rapamycin (mTOR). In recent years, some new mechanisms related to neuroinflammation have been shown to also play an important role in Mn-induced neurotoxicity. For example, DNA-sensing receptor cyclic GMP-AMP synthase (cCAS) and its downstream signal efficient interferon gene stimulator (STING), NOD-like receptor family pyrin domain containing 3(NLRP3)-pro-caspase1, cleaves to the active form capase1 (CASP1), nuclear factor κB (NF-κB), sirtuin (SIRT), and Janus kinase (JAK) and signal transducers and activators of the transcription (STAT) signaling pathway. Moreover, autophagy, as an important downstream protein degradation pathway, determines the fate of neurons and is regulated by these upstream signals. Interestingly, the role of autophagy in Mn-induced neurotoxicity is bidirectional. This review summarizes the molecular signaling pathways of Mn-induced neurotoxicity, providing insight for further understanding of the mechanisms of Mn.

Hair Follicle Development of Rex Rabbits Is Regulated Seasonally by Wnt10b/β-Catenin, TGFβ-BMP, IGF1, and EGF Signaling Pathways

This experiment was conducted to study the effects of different skinning seasons on the fur quality and hair follicle development of Rex rabbits. A total of 80,150-day-old Rex rabbits were slaughtered on 15 July 2022 (summer), 15 October 2022 (autumn), 15 January 2023 (winter), and 15 April 2023 (spring) in Shandong Province (10 males and 10 females in each season). The results show that the skin weight, skin area, skin thickness, and hair follicle density of the Rex rabbits (at 150 days of age) were lower in summer than in winter (p < 0.05). Moreover, the coat length was shorter in summer than in spring, autumn, and winter (p < 0.05). The shoulder fat weight, perirenal fat weight, and perigastric fat weight of the Rex rabbits in winter were higher than those in summer (p < 0.05). Furthermore, the leptin levels in serum were higher in winter than in summer in the Rex rabbits (p < 0.05). In terms of serum biochemistry, the glucose levels were higher in autumn and winter than in spring and summer (p < 0.05). The cholesterol, high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL) in summer had higher values than in winter in the Rex rabbits (p < 0.05). In winter, the expression of the Wnt10b, catenin beta 1 (CTNNB1), glycogen synthase kinase 3 beta (GSK3β), insulin like growth factor I (IGF-I), Type I insulin-like growth factor receptor (IGF-IR), and epidermal growth factor (EGF) genes was higher (p < 0.05), and the expression of the dickkopf-1 (DDK1), transforming growth factor beta 1 (TGFβ-1), bone morphogenetic protein 2 (BMP2), and bone morphogenetic protein 4 (BMP4) genes was lower than in summer (p < 0.05). In summer, the heat shock 70 kDa protein (HSP70) expression and CTNNB1 protein phosphorylation levels in skin tissue were higher than in spring, autumn, and winter (p < 0.05). In winter, Wnt10b protein expression was higher (p < 0.05), and GSK-3β protein phosphorylation levels were lower than in spring, autumn, and winter (p < 0.05). These results show that the skinning season can affect the production performance and hair follicle development of Rex rabbits. Compared with other seasons, the quality of skin from rabbits slaughtered in winter is better. Seasons may regulate hair follicle development via the Wnt10b/β-catenin, TGFβ-BMP, IGF1, and EGF signaling pathways in Rex rabbits.

Head organizer: Cerberus and IGF cooperate in brain induction in Xenopus embryos

Neural induction by cell-cell signaling was discovered a century ago by the organizer transplantations of Spemann and Mangold in amphibians. Spemann later found that early dorsal blastopore lips induced heads and late organizers trunk-tail structures. Identifying region-specific organizer signals has been a driving force in the progress of animal biology. Head induction in the absence of trunk is designated archencephalic differentiation. Two specific head inducers, Cerberus and Insulin-like growth factors (IGFs), that induce archencephalic brain but not trunk-tail structures have been described previously. However, whether these two signals interact with each other had not been studied to date and was the purpose of the present investigation. It was found that Cerberus, a multivalent growth factor antagonist that inhibits Nodal, BMP and Wnt signals, strongly cooperated with IGF2, a growth factor that provides a positive signal through tyrosine kinase IGF receptors that activate MAPK and other pathways. The ectopic archencephalic structures induced by the combination of Cerberus and IGF2 are of higher frequency and larger than either one alone. They contain brain, a cyclopic eye and multiple olfactory placodes, without trace of trunk structures such as notochord or somites. A dominant-negative secreted IGF receptor 1 blocked Cerberus activity, indicating that endogenous IGF signals are required for ectopic brain formation. In a sensitized embryonic system, in which embryos were depleted of β-catenin, IGF2 did not by itself induce neural tissue while in combination with Cerberus it greatly enhanced formation of circular brain structures expressing the anterior markers Otx2 and Rx2a, but not spinal cord or notochord markers. The main conclusion of this work is that IGF provides a positive signal initially uniformly expressed throughout the embryo that potentiates the effect of an organizer-specific negative signal mediated by Cerberus. The results are discussed in the context of the history of neural induction.

Abstract A150: Effects of radioimmunotherapy on human and canine osteosarcoma microenvironment

Abstract Introduction: Osteosarcoma (OS) is a type of primary bone cancer that predominantly affects children and adolescents with an unacceptably high mortality rate of 30% within 5 years of diagnosis. Though several treatment options including radiotherapy, chemotherapy, and amputation surgery exist, these are either ineffective or very invasive. Also, no significant advancements in treatment options have occurred in the last 30 years. The incidence of OS among companion dogs is much higher than in humans and phenotypically OS is very similar between humans and canines. We are using a comparative oncology approach to OS treatment by developing radioimmunotherapy (RIT) which targets the insulin-like growth factor receptor type 2 (IGF2R) on OS cells. IGF2R is abundant in human and canine OS and has shown promise a novel target for RIT of OS using a comparative oncology approach. It is therefore important to investigate the mechanistic underpinnings of RIT efficacy by investigating its interaction with OS microenvironment to develop improved RIT regimens for OS. Materials and Methods: Severe combined immunodeficiency (SCID) female mice bearing human and canine patient-derived OS xenografts were treated with RIT utilizing IF3, a novel full-length human mAb targeting IGF2R obtained by phage-display technology. The IF3 antibody was radiolabeled with an alpha emitter 225Actinium (225Ac) or a beta emitter 177Lutetim (177Lu) radionuclides. The tumors were removed at 1, 3 and 7 days post-RIT and the effects of RIT on IGF2R-positive tumor cells, OS stem cells, and immune components of the tumor microenvironment such as natural killer (NK) cells and tumor-associated macrophages (TAMs) were assessed using quantitative immunohistochemistry (IHC). Results: In the untreated tumors IHC detected the presence of IGF2R-positive tumor cells, OS stem cells, and immune components of the tumor microenvironment such as NK cells and TAMs, in both human and canine OS tumors obtained from mouse models. Quantitative analysis of IHC results in the tumors post-RIT demonstrated a time-dependent decrease in IGF2R-positive OS tumor cells and OS stem cells and an increase in TAMs and NK cells. Conclusions: Deciphering the mechanisms of RIT interaction with tumor microenvironment can assist in the design of the clinical trials in companion dogs with OS, which will be followed by the trials in children with this cancer. Also, it supports combining RIT with immunotherapy targeting OS stem cells and other immune components of tumor microenvironments such as NK cells and TAMS, which can result in improved patient outcomes. Citation Format: Sabeena Giri, Kevin J.H. Allen, Maruti Uppalapati, Ekaterina Dadachova. Effects of radioimmunotherapy on human and canine osteosarcoma microenvironment [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A150.

Insulin promotes growth in breast cancer cells through the type I IGF receptor in insulin receptor deficient cells

Breast cancer is the most common cancer in women. The upregulation of insulin-like growth factor (IGF) system observed in certain types of breast cancers was linked to growth, metastasis, and survival resulting in multiple strategies designed to target the type I IGF receptor (IGF-1R) in breast cancer. These attempts failed to prove beneficial and it has been suggested that insulin receptor (IR) could also play an important role in breast cancer biology. To better understand the IR's role in breast cancer cells, the receptor was deleted from MCF-7L cells using CRISPR technology, and fluorescence-assisted cell sorting was used to obtain clone 35 (CL35). It was found that CL35 activated signaling pathways upon insulin stimulation despite the absence of IR expression. We hypothesized that CL35 used a surrogate receptor for sustained growth and development. IGF-1R was able to activate insulin signaling and growth in CL35. Thus, insulin may play a central role in regulating breast cancer growth due to its ability to activate all the receptors of the IGF family. These findings argue that dual targeting of IR and IGF-IR may be required to inhibit breast cancer growth.

Klotho inhibits IGF1R/PI3K/AKT signalling pathway and protects the heart from oxidative stress during ischemia/reperfusion injury

Ischemia/reperfusion injury (IRI) of the heart involves the activation of oxidative and proapoptotic pathways. Simultaneously Klotho protein presents anti-aging, antiapoptotic and antioxidative properties. Therefore, this study aimed to evaluate the effect of Klotho protein on oxidative stress in hearts subjected to IRI. Isolated rat hearts perfused with the Langendorff method were subjected to ischemia, followed by reperfusion, in the presence or absence of recombinant rat Klotho protein. The factors involved in the activation of insulin-like growth factor receptor (IGF1R)/phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT) signalling pathway were evaluated. IRI caused activation of the IGF1R (p = 0.0122)/PI3K (p = 0.0022) signalling, as compared to the aerobic control group. Infusion supply of Klotho protein during IRI significantly reduced the level of phospho-IGF1R (p = 0.0436), PI3K (p = 0.0218) and phospho-AKT (p = 0.0020). Transcriptional activity of forkhead box protein O3 (FOXO3) was reduced (p = 0.0207) in hearts subjected to IRI, compared to aerobic control. Administration of Klotho decreased phosphorylation of FOXO3 (p = 0.0355), and enhanced activity of glutathione peroxidase (p = 0.0452) and superoxide dismutase (p = 0.0060) in IRI + Klotho group. The levels of reactive oxygen/nitrogen species (ROS/RNS) (p = 0.0480) and hydrogen peroxide (H2O2) (p = 0.0460), and heart injury (p = 0.0005) were significantly increased in hearts from the IRI group in comparison to the aerobic group. Klotho reduced NADPH oxidase 2 (NOX2) (p = 0.0390), ROS/RNS (p = 0.0435) and H2O2 (p = 0.0392) levels, and heart damage (p = 0.0286) in the hearts subjected to IRI. In conclusion, Klotho contributed to the protection of the heart against IRI and oxidative stress via inhibition of the IGF1R/PI3K/AKT pathway, thus can be recognized as a novel cardiopreventive/cardioprotective agent.

Mechanism of action of Huangbaichen Sanwei formulation in treating T2DM based on network pharmacology and molecular docking.

Huangbaichen Sanwei formulation (HBCS) has been reported to have a good hypoglycemic effect, but its pharmacological mechanism of action remains unclear. We used network pharmacology and molecular docking to explore the potential mechanism of action of HBCS against type-2 diabetes mellitus (T2DM). Fifty-five active components from HBCS interfered with T2DM. Twenty-five core targets, such as AKT1, INS, INSR, MAPK1 were identified. Enrichment analyses showed that HBCS was involved mainly including insulin receptor signaling pathway, extracellular region, and insulin-like growth factor receptor binding and other biological processes; common targets had roles in treating T2DM by regulating diabetic cardiomyopathy and insulin resistance. Molecular docking verified that components combined with core targets. HBCS play a part in treating T2DM through multiple components and targets at the molecular level, which lays a theoretical foundation for research using HBCS to treat T2DM. The components, predicted targets, and T2DM targets of HBCS were searched through databases, and common targets were determined. Further screening of the core targets was conducted through the establishment of a protein -protein interaction network. The core targets were analyzed by Gene Ontology (GO) annotation utilizing the DAVID platform. And the enrichment of signaling pathways was explored by employing the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Cytoscape 3.9.1 was employed to construct a "TCM-components-core target-pathway" network. Autodock Vina was used to dock molecules to compare the binding activity of active molecules with targets.

Molecular and serological biomarkers to predict trastuzumab responsiveness in HER-2 positive breast cancer.

HER-2-positive breast cancer is characterized by its aggressive nature, poor prognosis, and reduced overall survival. The emergence of trastuzumab resistance is currently considered a global problem. The immune system plays a pivotal role in tumor progression and development. Cytotoxic T lymphocyte-associated protein-4 (CTLA-4) and other immune checkpoint proteins may be potential prognostic factors and therapeutic targets for breast cancer. This study aimed to determine the correlation between CTLA-4 expression in peripheral blood and insulin-like growth factor-1 (IGF-1) serum levels and their impact on trastuzumab responsiveness in HER-2-positive patients with breast cancer. CTLA-4 expression was analyzed in peripheral blood cells using quantitative PCR, while IGF-1 serum levels were assessed through electrochemiluminescence assays. There was a significant increase in CTLA-4 expression at cycle 9, which continued to increase until it reached 4.6 at cycle 17. High IGF-1 levels were observed in newly diagnosed HER-2 positive patients before trastuzumab therapy, significantly decreasing post-therapy (p=0.001). Co-targeting HER-2 and IGF-1 receptors may reduce the risk of recurrence and improve outcomes. In addition, targeted CTLA-4 molecules may improve patient survival and prevent recurrence.

Improvement of oocyte competence and in vitro oocyte maturation with EGF and IGF-I in Guinea pig model

In vitro maturation (IVM) system is an alternative method to superovulation protocols to obtain mature oocytes. Epidermal Growth Factor (EGF) and Insulin-like Growth Factor I (IGF-I) have been widely used in IVM medium in different species. Although the guinea pig is a valuable animal model for reproductive studies, IVM is rarely used. We aimed to establish a suitable in vitro production system using EGF and/or IGF-I during IVM to improve oocyte competence. Firstly, immunolocalization of EGF and IGF-I receptors in the ovary was assessed. An IVM dose-response experiment was performed with cumulus-oocyte complexes (COCs) supplemented with: 1) EGF [0, 10, 50, 100 ng/mL or 10% fetal calf serum (FCS)]; 2) IGF-I [0, 50, 100, 200 ng/mL or 10% FCS]; or 3) the concentrations of EGF and IGF-I which showed the best IVM index in the previous experiments, with or without Fetal Calf Serum (FCS). Cortical granule and mitochondria distribution patterns were determined in in vivo and in vitro-matured oocytes for the first time in this species. Apoptotic rate after IVM and oocyte competence by in vitro embryo development were evaluated. Immunohistochemistry results showed positive immunostaining of EGF and IGF receptors in corpus luteum, oocytes, granulosa and theca cells in follicles in all stages of development. Supplementation of IVM medium with 50 ng/mL EGF or 100 ng/mL IGF-I or their combination with FCS successfully led to oocyte nuclear and cytoplasmic maturation and reduced the apoptotic rate. Both growth factors improved oocyte competence during IVM in this species since early embryos were in vitro developed, showing better results when FCS was used in the IVM medium.

Role of Insulin Like Growth Factor (IGF) in Lung Cancer with Underlying Diabetes Melitus

Lung cancer is one of the diseases which the highest incidence and mortality in the world. Lung cancer consists of small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) with various subtypes. Diabetes mellitus is one of the comorbidities that is often found in lung cancer patients, which is around 6% - 18% in lung cancer patients. Diabetes is a comorbidity factor that has a serious effect in increasing cancer cell progressivity through the activity of insulin growth factor (IGF) which consists of receptors (IGF-R) and binding proteins (IGFP). The role of IGF produces oncogenic mutations at the DNA level with the process of lung cancer immunity stimulates mitogenesis initiated by insulin and initiates neoplasm cells. IGF receptor mechanism has antineoplastic properties, to fulfill glycemia control that will reduce lung cancer progressivity and prevent clinical worsening so that it can help reduce lung cancer mortality with oral hypoglycemia drugs along with lung cancer therapy modalities such as cytotoxic agents, radiotherapy, immunotherapy, and surgery. The role of IGF is to amplify insulin levels in glucose, lipid, and protein metabolism. IGF can induce cell differentiation and initiation of neoplasm. Diabetes mellitus as a comorbidity can poor clinical outcomes and promote progression through IGF receptor activation in the tumorigenesis process.

Insulin Receptor Isoforms and Insulin Growth Factor-like Receptors: Implications in Cell Signaling, Carcinogenesis, and Chemoresistance.

This comprehensive review thoroughly explores the intricate involvement of insulin receptor (IR) isoforms and insulin-like growth factor receptors (IGFRs) in the context of the insulin and insulin-like growth factor (IGF) signaling (IIS) pathway. This elaborate system encompasses ligands, receptors, and binding proteins, giving rise to a wide array of functions, including aspects such as carcinogenesis and chemoresistance. Detailed genetic analysis of IR and IGFR structures highlights their distinct isoforms, which arise from alternative splicing and exhibit diverse affinities for ligands. Notably, the overexpression of the IR-A isoform is linked to cancer stemness, tumor development, and resistance to targeted therapies. Similarly, elevated IGFR expression accelerates tumor progression and fosters chemoresistance. The review underscores the intricate interplay between IRs and IGFRs, contributing to resistance against anti-IGFR drugs. Consequently, the dual targeting of both receptors could present a more effective strategy for surmounting chemoresistance. To conclude, this review brings to light the pivotal roles played by IRs and IGFRs in cellular signaling, carcinogenesis, and therapy resistance. By precisely modulating these receptors and their complex signaling pathways, the potential emerges for developing enhanced anti-cancer interventions, ultimately leading to improved patient outcomes.

THU543 Acquired Resistance To CDK4/6 Inhibitors Is Associated With Upregulated IGF1R And IR Signaling

Abstract Disclosure: S.L. Gerhardt: None. C. Baar: None. I. Johnson: None. D. Sachdev: None. The most common subtype of breast cancer is hormone receptor-positive (HR+). While these patients are successfully treated with endocrine therapies (ET), over 30% of patients develop resistance and require other therapies. Cyclin-dependent kinases (CDKs) 4 and 6 are required for cell cycle progression. Three CDK4/6 inhibitors (CDK4/6i), palbociclib, abemaciclib, and ribociclib, have recently been approved in combination with ET. CDK4/6i block phosphorylation of retinoblastoma (Rb) and cause G1/G0 arrest and are standard of care for treating metastatic HR+ Her2- advanced breast cancer with ET. Patients who respond to the CDK4/6i eventually develop resistance to the drugs. Thus, understanding the mechanisms of resistance to CDK4/6i and the role of growth-factor signaling will allow the identification of other pathways that can be targeted. To understand the mechanisms of resistance, we generated HR+ breast cancer cells (MCF-7) with acquired resistance to palbociclib (MCF-7/PalboR) or abemaciclib (MCF-7/AbemaR) by culturing them in increasing concentrations of the drugs. As shown by others, we found that acquired resistance to CDK4/6i results in loss of Rb and thus Rb phosphorylation. We analyzed signaling pathways and in vitro proliferation in the MCF-7/PalboR and MCF-7/AbemaR compared to the CDK4/6i sensitive matched parent cells (MCF-7L MP). MCF-7/PalboR and MCF-7/AbemaR had increased levels of CDK2 and cyclin E2. Both MCF-7/PalboR and MCF-7/AbemaR cells had upregulated type I IGF receptor (IGF1R) and insulin receptor (IR) protein levels compared MCF-7L MP as assayed by western blotting and flow cytometry. MCF-7/PalboR and MCF-7/AbemaR showed higher transcript levels of the fetal form of IR (IR-A) which is not expressed in normal adult tissues. IR-A plays a role in cancer progression and has been implicated in resistance to ET. MCF-7/PalboR and MCF-7/AbemaR had increased sensitivity to signaling by insulin-like growth factor I (IGF-I) and insulin compared to MCF-7 MP, with much lower concentrations of IGF-I and insulin phosphorylating the receptors and activating downstream MAPK and PI3K/Akt/mTOR pathways. Further, IGF-I and insulin mediated growth was enhanced to a statistically greater extent in MCF-7/PalboR and MCF-7/AbemaR compared to MCF-7 MP, and this enhanced growth was not blocked by 1200 nM of palbociclib or abemaciclib. MCF-7/PalboR cells were cross resistant to abemaciclib and MCF-7/AbemaR to palbociclib. Combined treatment with CDK4/6i and rapamycin, an mTOR pathway inhibitor, was not sufficient to overcome growth-factor mediated proliferation in MCF-7/PalboR and MCF-7/AbemaR but was in MCF-7 MP. These data indicate that acquired resistance to CDK4/6i is mediated through upregulated IGF1R and IR signaling. Our data suggest that combining CDK4/6i with drugs targeting IGF1R, IR-A, and mTOR signaling should be tested to delay resistance to CDK4/6i. Presentation: Thursday, June 15, 2023

Clinico-pathological characteristics of IGFR1 and VEGF-A co-expression in early and locally advanced-stage lung adenocarcinoma.

Although targeted therapies and immunotherapy have achieved significant clinical benefits in patients with certain pathological types of lung cancer. However, prognosis for patients with lung adenocarcinoma still remains unsatisfactory. It is of extremely importance to find ideal prognostic indicators to predict the prognosis of lung adenocarcinoma patients, especially for patients with early and locally advanced-stage lung adenocarcinoma. The purpose of this study is to elucidate the significance of Insulin-like growth factor receptor 1 (IGFR1) and Vascular endothelial growth factor A (VEGF-A) expression in predicting progression-free survival (PFS) and overall survival (OS) in patients with early and locally advanced-stage lung adenocarcinoma. In this study, IGFR1 and VEGF-A expression on 119 specimens of patients early and locally advanced-stage lung adenocarcinoma were analyzed by immunohistochemistry with an H-score system. Both high IGFR1 expression and VEGF-A expression patients were resulted in 59 (49.6%) separately. The numbers and proportions of IGFR1-&VEGF-A- subgroup, IGFR1-&VEGF-A+ subgroup, IGFR1+&VEGF-A- subgroup and IGFR1+&VEGF-A+ subgroup are 23 (19.3%), 37 (31.1%), 37 (31.1%) and 22 (18.5%) respectively. High IGFR1 expression was significantly associated with both poor PFS and OS of all patients in a univariate analysis. Multivariable analysis showed that patients with IGFR1+&VEGF-A+ expression exhibited a worst PFS and OS in the subgroup of lung adenocarcinoma patients with EGFR mutation. These results suggest that IGFR1+&VEGF-A+ is expected to be a disadvantageous factor for prognosis in the subgroup of EGFR mutation in patients with early and locally advanced-stage lung adenocarcinoma. What's more, this study may provide the theoretical possibility to screen optimal population for a combination therapy with anti-VEGF and anti-IGFR1 in patients with early and locally advanced-stage lung adenocarcinoma.