Macrophage Inhibitory Cytokine-1 Levels Research Articles

136 (PB126) - ALRN 6924 induces cell cycle arrest in bone marrow stem cells and hair follicles with dose-dependent degree and duration of effects after a single infusion in healthy volunteers

Background: ALRN-6924 is a cell-permeating, stabilized alpha-helical peptide that binds to endogenous p53 inhibitors MDMX and MDM2, thereby activating p53 and its transcriptional target p21 to cause cell cycle arrest (CCA). This effect is limited to cells with wild-type, functional p53. For cancer patients with tumors bearing mutated p53, ALRN-6924 treatment selectively induces CCA in normal cells, allowing chemotherapy to preferentially target p53-mutant cancer cells that are actively cycling. Materials and methods: A Phase 1 study in heathy volunteers is being conducted to evaluate ALRN-6924 pharmacokinetics (PK) and pharmacodynamics (PD). In Part 1 (37 subjects; Voors-Pette et al, ESMO 2021) it was shown that a 1-hour intravenous (IV) ALRN-6924 infusion was safe, well tolerated, and transiently upregulates p21 in human bone marrow (BM) cells with minimal signal for apoptosis. In Part 2, we aim to directly measure dose-dependent CCA in the BM as well as in hair follicles (HFs) in female subjects across a range of doses by 1-hour IV infusion and 3-minute bolus IV injection. ALRN-6924 was administered as a single IV infusion or bolus injection at 0 (placebo), 0.3, 0.6, or 0.9 mg/kg to cohorts of 3 to 5 subjects. Subjects were evaluated for safety and tolerability. Plasma and serum samples were obtained to determine PK and levels of macrophage inhibitory cytokine-1 (MIC-1), a biomarker of p53 activation. BM was sampled 12 hours post-dose to directly measure CCA by flow cytometry in CD34+, lineage-negative BM stem cells. Occipital scalp skin was sampled by 2 mm punch biopsy for p21 immunohistochemistry in HFs. Results: As of June 20, 2022, 42 subjects (ages 18–59; 100% female) were enrolled and evaluated in Part 2. Subjects experienced only mild, transient adverse events (AEs), with nausea/vomiting as the most frequent related AEs. The degree and duration of serum MIC-1 elevation was dose-dependent. At 12 hours post-dose, the proportion of cycling BM stem cells was significantly reduced at all dose levels, while blinded pathology review suggested ALRN-6924-dependent p21 induction in anagen-phase HFs. Safety profiles, PK and PD were similar for both bolus and infusion. Conclusions: ALRN-6924 shows a favorable safety profile for use as a chemoprotection agent. CCA measured in BM supports prevention of chemotherapy-induced neutropenia, thrombocytopenia, and anemia, while HF results support chemoprotection for alopecia. Administration by IV bolus may simplify dosing vs. the current 1-hr infusion clinical regimen. The dose-dependent degree and duration of effects support development of a universal treatment schedule for broad use across cancer indications and types of chemotherapy. Conflict of interest: Ownership: Manuel Aivado, Vojislav Vukovic, and Allen Annis are employees and shareholders of Aileron Therapeutics Advisory Board: Ulrich Steidl is a member of the Scientific Advisory Board of Aileron Therapeutics Other Substantive Relationships: Britta Will and Kevin Randall are consultants to Aileron Therapeutics

Clinical usefulness and acceleratory effect of macrophage inhibitory cytokine-1 on biliary tract cancer: an experimental biomarker analysis

BackgroundBiliary tract cancer (BTC) has a poor prognosis; therefore, useful biomarkers and treatments are needed. Serum levels of macrophage inhibitory cytokine-1 (MIC-1), a member of the TGF-β superfamily, are elevated in patients with pancreaticobiliary cancers. However, the effect of MIC-1 on BTC is unknown. Therefore, we investigated the effect of MIC-1 on BTC and assessed whether MIC-1 is a biomarker of or therapeutic target for BTC.MethodsMIC-1 expression in BTC cells was determined by performing histological immunostaining, tissue microarray (TMA), western blotting, and reverse transcription PCR (RT–PCR). Cell culture experiments were performed to investigate the effect of MIC-1 on BTC cell lines (HuCCT-1 and TFK-1). The relationships between serum MIC-1 levels and either the disease state or the serum level of the apoptosis marker M30 were retrospectively verified in 118 patients with pancreaticobiliary disease (individuals with benign disease served as a control group, n = 62; BTC, n = 56). The most efficient diagnostic marker for BTC was also investigated.ResultsMIC-1 expression was confirmed in BTC tissue specimens and was higher in BTC cells than in normal bile duct epithelial cells, as determined using TMA, western blotting and RT–PCR. In cell culture experiments, MIC-1 increased BTC cell proliferation and invasion by preventing apoptosis and inhibited the effect of gemcitabine. In serum analyses, serum MIC-1 levels showed a positive correlation with BTC progression and serum M30 levels. The ability to diagnose BTC at an early stage or at all stages was improved using the combination of MIC-1 and M30. The overall survival was significantly longer in BTC patients with serum MIC-1 < the median than in BTC patients with serum MIC-1 ≥ the median.ConclusionsMIC-1 is a useful diagnostic and prognostic biomarker and might be a potential therapeutic target for BTC.

Macrophage inhibitory cytokine-1 produced by melanoma cells contributes to melanoma tumor growth and metastasis in vivo by enhancing tumor vascularization.

Macrophage inhibitory cytokine-1 (MIC-1) has been reported to be elevated in various human cancers including melanoma; however, the function of MIC-1 in cancer remains unclear. In this study, we attempt to clarify the role of MIC-1 in tumor pathogenesis by employing the orthotopic B16F1 melanoma mouse model in which serum MIC-1 levels are positively correlated with tumor size. By stably transfecting a MIC-1 expression construct into B16F1 melanoma cells, we increased the expression and secretion levels of MIC-1. This increase in MIC-1 expression significantly enhanced the growth of tumors derived from B16F1 cells in vivo, despite not affecting in vitro cell growth. The elevated MIC-1 expression in B16F1 cells also resulted in lymph node metastasis in B16F1 tumor-bearing mice, significantly increasing mortality. Interestingly, among small melanoma tumors of similar size, tumors derived from the MIC-1-transfected B16F1 cells exhibited enhanced blood vessel formation compared with those of mock transfectant cells. Also, more MIC-1 was found in well-vascularized tumor regions than in poorly vascularized tumor regions. Moreover, conditioned medium (CM) of the MIC-1-transfected melanoma cells enhanced the angiogenic properties of endothelial cells more than CM of mock transfectant cells. Notably, hypoxic culture conditions forced parental B16F1 cells to secrete more endothelial cell-stimulating factors, among which the function of MIC-1 was confirmed by blocking the effects with an anti-MIC-1 antibody. Taken together, these results suggest that the MIC-1 produced by melanoma cells in response to oxygen deprivation promotes tumor vascularization during melanoma development in vivo, leading to enhanced tumor growth and metastasis.

Abstract P211: The investigational chemoprotection drug ALRN-6924, a dual inhibitor of MDMX and MDM2, shows potential for radioprotection

Abstract Aim: We investigated whether p53 activation with ALRN-6924 can prevent toxicity in mouse models of acute radiation injury. Materials and methods: ALRN-6924 is a clinical-stage, first-in-class, stabilized cell-permeating alpha-helical peptide that disrupts the interaction of the p53 tumor suppressor protein with its endogenous inhibitors, MDMX and MDM2. In previous experiments it was shown that ALRN-6924 can induce transient, dose-dependent cell cycle arrest in the bone marrow to protect those cells from chemotherapy while not protecting p53-mutant cancer cells. Because radiation (like chemotherapy) preferentially affects proliferating cells, we hypothesized that ALRN-6924 may also protect proliferating cells in normal tissues from radiation-induced cellular toxicity. Cell cycle arrest was measured in the bone marrow of ALRN-6924-treated C57BL/6 mice by flow cytometry using EdU incorporation. Serum levels of macrophage inhibitory cytokine-1 (MIC-1), a biomarker of p53 activation, were measured by ELISA. Cell proliferation and apoptosis were measured in formalin-fixed mouse bone marrow by immunohistochemistry analysis of Ki67 and cleaved PARP, respectively. C57BL/6 mice (n=7/group) were treated with one or more intravenous 2.4 mg/kg doses of ALRN-6924 at 24, 16, 8, or 1 hour (or combinations thereof) or placebo prior to an abdominally targeted (shielded body) 15 Gy radiation dose and then monitored for body weight (BW). Results: MIC-1 was elevated in the serum of ALRN-6924-treated mice in a dose-dependent fashion. Repeated doses of ALRN-6924 every 8 hrs yielded sustained MIC-1 elevation, which correlated with reduced Ki67 positivity in the bone marrow. Treatment-dependent changes in cPARP expression were evident, but minimal in magnitude. In a nonlethal radiation exposure model, ALRN-6924 yielded significant protection from radiation-induced BW loss in a schedule-dependent manner. Placebo-treated mice showed 10% to 15% BW loss five days after irradiation, while mice receiving one or more ALRN-6924 doses 8 hrs prior to irradiation had an average of 4% BW loss (p=0.008, two-sided t test). Conclusions: ALRN-6924 mitigates toxicity in a mouse model of acute radiation injury. The observed radioprotection effect correlates with pharmacodynamic markers of cell proliferation and cell cycle arrest after one or more doses of ALRN-6924, and further supports previous demonstrations of chemoprotection with ALRN-6924. These results provide a rationale to investigate ALRN-6924 as a radioprotective agent. Citation Format: Allen Annis, David Sutton, Manuel Aivado, Vojislav Vukovic. The investigational chemoprotection drug ALRN-6924, a dual inhibitor of MDMX and MDM2, shows potential for radioprotection [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P211.

Alrn-6924, a Dual Inhibitor of MDMX and MDM2, Transiently Induces Cell Cycle Arrest in Bone Marrow and Prevents Toxicity in Mouse Models of Acute Radiation Injury

Background: Myelosuppression is a common sequela of acute radiation injury due to sensitivity of proliferating bone marrow cells to ionizing radiation. ALRN-6924 is a clinical-stage, first-in-class, stabilized cell-permeating alpha-helical peptide drug that disrupts the interaction of the p53 tumor suppressor protein with its endogenous inhibitors, MDMX and MDM2, to induce transient, dose-dependent cell cycle arrest in p53-wild-type tissues. ALRN-6924 is being evaluated in clinical trials as a selective chemoprotection agent for patients with p53-mutant cancers to protect healthy normal cells from chemotherapy while not protecting p53-mutant cancer cells. We tested whether ALRN-6924 may similarly protect against radiation-induced toxicity in mouse models of acute radiation injury.Materials and methods: Activation of p21 (CDKN1A), a cell cycle regulator under transcriptional control of p53, was measured in formalin-fixed mouse bone marrow by immunohistochemistry analysis (IHC). Proliferation and apoptosis in bone marrow were measured by IHC of Ki67 and cleaved PARP, respectively. Cell cycle arrest was measured in the bone marrow of ALRN-6924-treated C57BL/6 mice by flow cytometry using EdU incorporation. Serum levels of macrophage inhibitory cytokine-1 (MIC-1), a biomarker of p53 activation, were measured by ELISA. As a model of radiation-induced toxicity, C57BL/6 mice (n=7/group) were treated with one or more intravenous 2.4 mg/kg doses of ALRN-6924 at 24, 16, 8, or 1 hour (or combinations thereof) or placebo prior to a 15 Gy shielded-body radiation dose and then monitored for body weight (BW) changes.Results: ALRN-6924 induced cell cycle arrest in mouse bone marrow with a maximum effect at 8 hrs post-dose. Repeated doses of ALRN-6924 every 8 hrs elevated p21 levels in bone marrow that correlated with reduced Ki67 positivity and increased serum MIC-1 levels. Treatment-dependent changes in cPARP expression in bone marrow were evident, but minimal in magnitude. In a nonlethal radiation exposure model, ALRN-6924 yielded significant protection from radiation-induced BW loss in a schedule-dependent manner. Placebo-treated mice showed 10% to 15% BW loss five days after irradiation, while mice receiving one or more ALRN-6924 doses 8 hrs prior to irradiation had an average of 4% BW loss (p=0.008, two-sided t test).Conclusions: ALRN-6924 mitigated toxicity in a mouse model of acute radiation injury. The observed radioprotection effect was correlated with cell cycle arrest in bone marrow after one or more doses of ALRN-6924. These results support further investigation of ALRN-6924 as a radioprotective agent. DisclosuresAnnis: Aileron Therapeutics, Inc.: Current Employment. Sutton: Aileron Therapeutics, Inc.: Consultancy; Kriya Therapeutics: Consultancy; First Light Pharmaceuticals: Consultancy; Cygnal Therapeutics: Consultancy. Aivado: Aileron Therapeutics, Inc.: Current Employment. Vukovic: Aileron Therapeutics, Inc.: Current Employment.

Circulatory MIC-1 as a Determinant of Prostate Cancer Racial Disparity.

Simple SummaryAfrican American men are diagnosed with more aggressive prostate cancer and have worse outcomes than Caucasians. This study examined the role of MIC-1 as a risk factor and demonstrated a conceptual observation for the differential level of MIC-1 in circulation (serum and urine) and tumor tissues from prostate cancer patients of racial disparity. The circulatory MIC-1 levels in serum and urine are significantly higher in prostate cancer patients of African American ethnicity, with higher sensitivity and specificity than Caucasians. The validation of circulatory MIC-1 in a larger cohort of patients may help identify high-risk prostate cancer patients and develop race-oriented therapies to reduce the observed cancer outcome gaps between the races.In this study, we investigated the potential of MIC-1 (macrophage inhibitory cytokine-1) on the severity of prostate cancer between African American men and Caucasians. Differences between the races were examined using Mann–Whitney tests for continuous variables and Fisher’s exact tests for categorical variables. Pearson’s correlation coefficient was used to identify associations between continuous measures across all samples and within each race. Analysis of variance, including clinical parameters, was used to identify differences in serum and urine MIC-1 levels between races. We found significant differences between the two races for age (p = 0.01), Gleason scores (p = 0.01), and stage of disease (p = 0.03). African American men in the study had higher Gleason scores (mean = 6.9) than Caucasians (mean = 6.5), during earlier stages of the disease. In Caucasian men with prostate cancer, serum MIC-1 expression was positively associated with age (r = 0.7, p < 0.01). However, African American men had highly expressed MIC-1 and high Gleason scores (r = 0.16, p = 0.3). Interestingly, the urine MIC-1 level was significantly higher in African American men with prostate cancer than in Caucasian patients. It appeared to be more sensitive and specific for African Americans (AUC = 0.85 vs. 0.56). Thus, high circulatory MIC-1 in prostate cancer patients may indicate MIC-1 as a potential biomarker to improve the diagnostic ability of an aggressive stage of prostate cancer in African American men. However, a larger cohort of sample analysis is required to validate these observations.

Plasma Macrophage Inhibitory Cytokine-1 as a Complement of Epstein-Barr Virus Related Markers in Identifying Nasopharyngeal Carcinoma.

Background:We evaluated the diagnostic value of plasma Macrophage inhibitory cytokine-1 (MIC-1) in distinguishing patients with nasopharyngeal carcinoma (NPC) and explored its complementary role with widely used Epstein-Barr virus (EBV) related markers, EBV capsid antigen-specific IgA (VCA-IgA) and EBV copy number.Methods:ELISA was used to analyze the plasma MIC-1 levels in 190 NPC patients, 72 VCA-IgA-positive healthy donors (VP), and 219 normal subjects with negative VCA-IgA (VN). 10 pairs of plasma samples before and after radiotherapy were also included.Results:The plasma MIC-1 levels were significantly higher in NPC patients (Median: 678.39 ng/mL) than those in VN and VP (310.29 and 294.59, p < 0.001). Receiver operating characteristic (ROC) curves of the MIC-1 concentrations revealed that the area under the ROC curve (AUC) was 0.790 (95% confidence interval [CI]: 0.748-0.832), with a sensitivity of 63.7%, and a specificity of 85.9% respectively, for distinguishing NPC patients from the healthy donors. Similarly, between NPC and VP, ROC was 0.796 (0.738-0.853) with sensitivity of 63.7%, and specificity of 88.9%. In addition, between NPC and VN, ROC was 0.788(0.744-0.832) with sensitivity of 63.7%, and specificity of 84.9%. Further, we found that MIC-1 could complement VCA-IgA and EBV DNA markers, with a negative rate of 88.9% in VCA-IgA-positive healthy controls, and a positive rate of 59.0% in EBV DNA negative NPC patients, respectively. Also, the MIC-1 plasma concentration dropped significantly after radiotherapy (p = 0.027).Conclusions:MIC-1 can complement VCA-IgA titers and EBV DNA copy number tests in NPC detection, improve identification of EBV DNA-negative NPC patients, and distinguish NPC from VCA -IgA positive healthy controls.

Clinical application of combined detection of serum macrophage inhibitory cytokine-1, carcinoembryonic antigen and carbohydrate antigens for the diagnosis and prognosis of colorectal cancer in the elderly

Objective To explore the application value of combined detection of serum macrophage inhibitory cytokine-1(MIC-1), carcinoembryonic antigen(CEA)and carbohydrate antigens in the diagnosis and prognosis of elderly patients with colorectal cancer.172 elderly patients with colorectal cancer were selected retrospectively as the observation group, 175 elderly patients with benign colorectal lesions were enrolled as the benign lesion group and 166 healthy elderly subjects as the control group. Methods The levels of MIC-1, CEA, carbohydrate antigens of 199(CA199), 724(CA724), 242(CA242), 125(CA125), and 50(CA50)were compared among the three groups, and the diagnostic efficacy was calculated.The difference in the above-mentioned indicators between patients with different stages and prognosis in the observation group were compared. Results Levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 were higher in the observation group than in the benign lesion group and the control group, and the above indexes were higher in the benign lesion group than in the control group.The sensitivity, positive predictive value and negative predictive value of MIC-1 were the highest in single index test(68.6%, 72.8% and 84.6%, respectively). The specificity of CA199 was the highest(91.2%). The combined detections greatly improved sensitivity, positive predictive value and negative predictive value(89.0%, 79.3% and 94.1%, respectively). In the observation group, levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 in the elderly colorectal cancer patients were higher in stage Ⅲ and Ⅳ than in stage Ⅰ and Ⅱ.The levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 in elderly patients with recurrence and metastasis of colorectal cancer were higher than those in patients without recurrence and metastasis. Conclusions Combined detection of multiple indicators can improve the sensitivity of diagnosis, and is more conducive to the early diagnosis of elderly patients with colorectal cancer. Key words: Colorectal neoplasms; Macrophage migration-inhibitory factors; Carcinoembryonic antigen; Antigens, tumor-associated, carbohydrate

Elevated circulating macrophage inhibitory cytokine 1 is a biological marker for the diagnosis and prognosis of osteosarcoma.

Osteosarcoma is among the most frequently occurring bone tumors in infants and teenagers. However, despite its widespread prevalence, no effective diagnostic and prognostic biomarkers for osteosarcoma are known. Macrophage inhibitory cytokine 1 (MIC-1) has been considered a promising biological marker of various tumor types. However, the possible role of circulating MIC-1 as a screening biomarker for osteosarcoma remains to be elucidated. The present study evaluated the circulating levels of MIC-1 in patients with osteosarcoma with the aim of elucidating its effect on the diagnosis and prognosis of this specific tumor. The circulating levels of MIC-1 were measured via an enzyme-linked immunosorbent assay in 300 individuals, including 100 patients with osteosarcoma, 100 patients with benign bone tumors, and 100 healthy subjects, and its correlation with clinicopathological characteristics was then evaluated. Various analyses were performed to determine its utility in diagnosis and prognosis. The levels of circulating MIC-1 were increased considerably in patients with osteosarcoma. The patients bearing larger tumors, those with distant metastases, and those with later-stage tumors had relatively higher levels of MIC-1. According to the multivariate logistic regression analysis, a high level of circulating MIC-1 was an independent variable for distant metastases. Receiver operating characteristic analysis revealed MIC-1 as a possible biological marker for distinguishing patients from healthy controls. Patients with osteosarcoma with higher levels of MIC-1 had relatively higher risk of mortality. Furthermore, multivariate data analysis on general survival rate revealed that a high level of circulating MIC-1 was a prognostic indicator of osteosarcoma. These findings suggest that an elevated level of circulating MIC-1 is a novel potential diagnostic and prognostic biomarker for osteosarcoma.

The value of macrophage inhibitory cytokine-1 level in differentiating benign from malignant solitary pulmonary nodules.

Macrophage inhibitory cytokine-1 (MIC-1), a transforming growth factor-β superfamily cytokine, is involved in tumor pathogenesis, and its measurement can be used as a clinical tool for the diagnosis of a wide range of cancers. The aim of this study was to explore the diagnostic value of serum MIC-1 in patients with solitary pulmonary nodules (SPNs). Serum specimens from 158 malignant SPN patients, 110 benign SPN patients, along with 120 healthy volunteers. The levels of serum MIC-1 were measured by sandwich enzyme-linked immunosorbent assay. Serum levels of MIC-1 in malignant SPN patients were significantly higher than those in benign SPN patients (P < .01), or those in healthy volunteers (P < .01). With a cutoff of 685.8 pg/ml, the sensitivity and specificity of MIC-1 in differentiating between malignant SPN patients and benign SPN patients, and between malignant SPN patients and healthy volunteers was, 56.3% and 92.7%, and 65.8% and 96.7%, respectively. An area under the curve (AUC) for malignant SPN resulting from MIC-1, which was significantly better than any other tumor markers tested including carbohydrate antigens 12-5 (CA125), and carcinoembryonic antigen (CEA). In conclusion, measurement of serum MIC-1 levels could be considered as a diagnostic biomarker for malignant SPN patients.

The efficacy of biliary and serum macrophage inhibitory cytokine-1 for diagnosing biliary tract cancer

The serum macrophage inhibitory cytokine-1 (MIC-1) levels are elevated in some inflammatory conditions and cancers. We thus compared the levels of biliary and serum MIC-1 and conventional tumour markers between 23 biliary tract cancer (BTC) patients (malignant group) and 29 benign biliary disease patients (benign group) and found that all markers were significantly elevated in the malignant group. The levels of two markers were higher in early BTC (Stage I/II, n = 15) than in the benign group: biliary MIC-1 [12 (0–2153) vs. 678 (0–4429) pg/ml, P < 0.01] and serum CA19–9 [13 (2–15,682) vs. 45.1 (2–10,478) U/ml, P = 0.02]. A receiver operating characteristic curve analysis revealed that the area under the curve for biliary MIC-1 was greater than that for serum CA19-9 (0.77 vs. 0.73). The cut-off value for biliary MIC-1 in diagnosing early BTC was 581.6 pg/ml, and this value yielded a sensitivity, specificity and accuracy of 71.4%, 82.8%, and 79.1%, respectively. The sensitivity of biliary MIC-1 for diagnosing early BTC was superior to that of biliary cytology (71.4% vs. 8.33%, P < 0.01), and the combination of serum MIC-1 with CA19-9 (cut-off value = 4021.2 pg/ml, 42.4 U/ml) was useful for screening BTC (sensitivity = 82.6%, specificity = 72.4%). In conclusion, biliary MIC-1 can effectively diagnose early BTC.

Expression of macrophage inhibitory cytokine-1 in early gastric cancer cases treated using endoscopic mucosal resection and the correlation with prognosis.

The expression of macrophage inhibitory cytokine-1 (MIC-1) of early gastric cancer treated using endoscopic mucosal resection (EMR) and its correlation with prognosis were investigated. One hundred and forty-seven patients with early gastric cancer (less than 20 mm in diameter) were recruited and assigned into an observation group (75 cases) or a control group (72 cases), according to treatment. The control group was treated with a radical resection by the conventional laparotomy approach and the observation group was treated by EMR. Differences in operative outcomes and prognosis were compared between the two groups. Complete lesion resection rates of the two groups were 100%. The amount of bleeding, operation time and postoperative complication rates in the observation group were each significantly lower than those in the control group (P<0.05). The serum MIC-1 levels in the two groups were reduced, with the observation group being significantly lower than that of the control group (P<0.05). The progression-free survival was significantly prolonged and the recurrence rate was significantly lower in the observation group compared to the control group, with the difference being statistically significant (P<0.05). In conclusion, EMR treatment of early gastric cancer was observed to be safe and effective in this study and can reduce the expression level of serum MIC-1 after the operation. EMR can also increase the survival period and reduce the recurrence rate, and MIC-1 can be used as an important index to predict the prognosis of the disease.

Oral Treatment with the Ghrelin Receptor Agonist HM01 Attenuates Cachexia in Mice Bearing Colon-26 (C26) Tumors.

The gastrointestinal hormone ghrelin reduces energy expenditure and stimulates food intake. Ghrelin analogs are a possible treatment against cancer anorexia-cachexia syndrome (CACS). This study aimed to investigate whether oral treatment with the non-peptidergic ghrelin receptor agonist HM01 counteracts CACS in colon-26 (C26) tumor-bearing mice. The C26 tumor model is characterized by pronounced body weight (BW) loss and muscle wasting in the absence of severe anorexia. We analyzed the time course of BW loss, body composition, muscle mass, bone mineral density, and the cytokines interleukin-6 (IL-6) and macrophage-inhibitory cytokine-1 (MIC-1). Moreover, we measured the expression of the muscle degradation markers muscle RING-finger-protein-1 (MuRF-1) and muscle atrophy F-box (MAFbx). After tumor inoculation, MIC-1 levels increased earlier than IL-6 and both cytokines were elevated before MuRF-1/MAFbx expression increased. Oral HM01 treatment increased BW, fat mass, and neuronal hypothalamic activity in healthy mice. In tumor-bearing mice, HM01 increased food intake, BW, fat mass, muscle mass, and bone mineral density while it decreased energy expenditure. These effects appeared to be independent of IL-6, MIC-1, MuRF-1 or MAFbx, which were not affected by HM01. Therefore, HM01 counteracts cachectic body weight loss under inflammatory conditions and is a promising compound for the treatment of cancer cachexia in the absence of severe anorexia.

First Behavioural Characterisation of a Knockout Mouse Model for the Transforming Growth Factor (TGF)-β Superfamily Cytokine, MIC-1/GDF15.

Macrophage inhibitory cytokine-1 (MIC-1), also known as growth differentiation factor 15 (GDF15), is a stress response cytokine. MIC-1/GDF15 is secreted into the cerebrospinal fluid and increased levels of MIC-1/GDF15 are associated with a variety of diseases including cognitive decline. Furthermore, Mic-1/Gdf15 knockout mice (Mic-1 KO) weigh more, have increased adiposity, associated with increased spontaneous food intake, and exhibit reduced basal energy expenditure and physical activity. The current study was designed to comprehensively determine the role of MIC-1/GDF15 on behavioural domains of male and female knockout mice including locomotion, exploration, anxiety, cognition, social behaviours, and sensorimotor gating. Mic-1 KO mice exhibited a task-dependent increase in locomotion and exploration and reduced anxiety-related behaviours across tests. Spatial working memory and social behaviours were not affected by Mic-1/Gdf15 deficiency. Interestingly, knockout mice formed an increased association with the conditioned stimulus in fear conditioning testing and also displayed significantly improved prepulse inhibition. Overall sex effects were evident for social behaviours, fear conditioning, and sensorimotor gating. This is the first study defining the role of Mic-1/Gdf15 in a number of behavioural domains. Whether the observed impact is based on direct actions of Mic-1/Gdf15 deficiency on the CNS or whether the behavioural effects are mediated by indirect actions on e.g. other neurotransmitter systems must be clarified in future studies.

Macrophage Inhibitory Cytokine-1 as a Novel Diagnostic and Prognostic Biomarker in Stage I and II Nonsmall Cell Lung Cancer.

Background:Increased level of serum macrophage inhibitory cytokine-1 (MIC-1), a member of transforming growth factor-β superfamily, was found in patients with epithelial tumors. This study aimed to evaluate whether serum level of MIC-1 can be a candidate diagnostic and prognostic indicator for early-stage nonsmall cell lung cancer (NSCLC).Methods:A prospective study enrolled 152 patients with Stage I–II NSCLC, who were followed up after surgical resection. Forty-eight patients with benign pulmonary disease (BPD) and 105 healthy controls were also included in the study. Serum MIC-1 levels were measured using an enzyme-linked immunosorbent assay, and the association with clinical and prognostic features was analyzed.Results:In patients with NSCLC, serum protein levels of MIC-1 were significantly increased compared with healthy controls and BPD patients (all P < 0.001). A threshold of 1000 pg/ml of MIC-1 was found in patients with early-stage (Stage I and II) NSCLC, with sensitivity and specificity of 70.4% and 99.0%, respectively. The serum levels of MIC-1 were associated with age (P = 0.001), gender (P = 0.030), and T stage (P = 0.022). Serum MIC-1 threshold of 1465 pg/ml was found in patients with poor early outcome, with sensitivity and specificity of 72.2% and 66.1%, respectively. The overall 3-year survival rate of NSCLC patients with high serum levels of MIC-1 (≥1465 pg/ml) was lower than that of NSCLC patients with low serum MIC-1 levels (77.6% vs. 94.8%). Multivariate Cox regression survival analysis showed that a high serum level of MIC-1 was an independent risk factor for reduced overall survival (hazard ratio = 3.37, 95% confidential interval: 1.09–10.42, P = 0.035).Conclusion:The present study suggested that serum MIC-1 may be a potential diagnostic and prognostic biomarker for patients with early-stage NSCLC.

Macrophage Inhibitory Cytokine-1 (MIC-1) as A Biomarker for Diagnosis and Prognosis of Stage I-II Non-small Cell Lung Cancer

背景与目的巨噬细胞抑制因子-1（macrophage inhibitory cytokine-1, MIC-1）是人转化生长因子β（transforming growth factor-β, TGF-β）超家族中重要成员，研究发现MIC-1表达水平在多种上皮来源肿瘤患者血清中均有显著升高。本研究旨在探讨MIC-1在早期非小细胞肺癌（non-small cell lung cancer, NSCLC）诊断及其与临床病理特征间的关系，以及与术后复发/转移及预后的相关性。方法采用酶联免疫吸附试验（enzymelinked immunosorbent assay, ELISA）方法检测152例早期肺癌、48例肺良性疾病患者及105例正常对照人群血清MIC-1浓度，分析MIC-1诊断肺癌中的作用，同时分析血清MIC-1浓度与临床病理特征、复发/转移及预后的相关性。结果早期肺癌患者组MIC-1血清水平高于正常对照组（P < 0.001）和肺良性疾病组（P < 0.001），设1, 000 pg/mL为诊断肺癌的临界值，MIC-1检测肺癌的敏感性和特异性分别为70.4%和99.0% [曲线下面积（area under curve, AUC）: 0.90；95%CI: 0.87-0.94]；MIC-1血清水平与年龄（P=0.001）、性别（P=0.03）有关，病理TNM分期T2的患者MIC-1血清水平高于T1患者（P=0.022）；血清MIC-1 > 1, 465 pg/mL组的患者3年生存率为77.6%，低于血清MIC-1 < 1, 465 pg/mL组的患者94.8%（P=0.022），Cox回归多因素分析结果显示，血清MIC-1 > 1, 465 pg/mL是Ⅰ期、Ⅱ期NSCLC独立的预后因素（HR=3.37, 95%CI: 1.09-10.42, P=0.035）。结论MIC-1作为血清肿瘤生物标志物，可能有助于提高肺癌早期诊断。MIC-1的检测对判断Ⅰ期、Ⅱ期NSCLC患者预后有预测价值，可能为其独立的预后指标。

112P Macrophage inhibitory cytokine-1 as a biomarker for diagnosis and prognosis of stage I-II non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death and has a poor prognosis. Biomarkers should be established to improve patient care especially in early-stage NSCLC. The macrophage inhibitory cytokine-1 (MIC-1) can promote tumor invasiveness and metastasis. Therefore, our aim was to delineate the diagnostic and prognostic value of serum MIC-1 in patients with early-stage NSCLC. A total of 152 consecutive patients with stage I–II NSCLC were retrospectively reviewed and underwent follow up after total resection of tumor. Serum MIC-1 level was evaluated in patients and 229 healthy controls, and was correlated with clinical features and prognosis of patients. The level of MIC-1 of NSCLC patients was significantly higher than that of controls (P < 0.001). A threshold of 1000 pg/ml could be used to diagnose early-stage NSCLC with 96.5% specificity and 71.3% sensitivity (area under curve, 0.912; 95% CI, 0.886–0.939). Higher level of MIC-1 was associated with elder age (> 60 years old) at diagnosis (P = 0.001) and female gender (P = 0.03). There was a significant difference between the overall survival for patients with high level of MIC-1 (≥1000 pg/ml) and low level of MIC-1(log rank, P = 0.045). The overall 3-year survival rate in patients with high level of MIC-1 was significantly lower than that of patients with low MIC-1 level (84.0% vs. 97.8%, P < 0.05). Cox regression revealed that a high level of MIC-1 was a potential risk factor (HR = 7.848, 95%CI, 1.044–58.979) for compromised overall survival. The high level of serum MIC-1 might be served as a potential biomarker for diagnosis and poorer outcome in patients with early-stage NSCLC. The clinical significance could be evaluated in future studies with larger sample size.

Plasma Inflammatory Markers and Risk of Advanced Colorectal Adenoma in Women

Evidence remains inconclusive about the association of systemic inflammatory markers with colorectal neoplasia. We investigated whether circulating inflammatory markers were associated with risk of advanced colorectal adenoma. We measured plasma macrophage inhibitory cytokine-1 (MIC-1), C-reactive protein (CRP), interleukin-6 (IL6), and soluble TNF receptor 2 (sTNFR-2) in blood samples drawn from 32,826 women in 1989 to 1990 in the Nurses' Health Study. Through 2008, we documented 757 cases of advanced colorectal adenomas (≥1 cm or any size with advanced histology); each case was matched by age and time of blood draw with one control randomly selected from participants who underwent lower endoscopy and did not have neoplasia. Plasma MIC-1 was associated with higher risk of advanced adenoma (Ptrend = 0.04), with an OR of 1.55 (95% confidence interval, 1.03-2.32) comparing extreme quintiles of MIC-1 after adjusting for colorectal cancer-risk factors and other inflammatory markers. Among cases, MIC-1 level was positively associated with the number of adenomas (P < 0.001) and gradually increased from adenomas located in the rectum, distal colon, and up to the proximal colon. There was a strong positive association between MIC-1 and risk of adenomas with multiplicity, ≥1 cm size and location in the proximal colon (all Ptrend < 0.05). CRP, IL6, or sTNFR-2 was not associated with adenoma risk. In conclusion, plasma MIC-1 was associated with higher risk of colorectal adenoma, especially multiple, large, and proximal adenomas. Our results provide further support for a role for MIC-1 in carcinogenesis and the potential for MIC-1 as an adjunctive biomarker for detection of advanced colorectal adenoma.

Association Between Plasma Levels of Macrophage Inhibitory Cytokine-1 Before Diagnosis of Colorectal Cancer and Mortality

Patients with colorectal cancer (CRC) have high circulating levels of macrophage inhibitory cytokine-1 (MIC1 or growth differentiation factor 15), a marker of inflammation that might be involved in carcinogenesis. We analyzed blood samples collected from individuals before they were diagnosed with CRC to determine whether levels of MIC1 were associated with mortality. We collected data on survival of 618 participants diagnosed with CRC who provided prediagnosis blood specimens in 1990 (Nurses' Health Study) and 1994 (Health Professionals' Follow-up Study) and were followed through 2010. Levels of MIC1 were measured by enzyme-linked immunosorbent assay and then were categorized into quartiles based on the known distribution of MIC1 levels among previously matched individuals without CRC (controls) within each cohort. We then examined the association of MIC1 levels with overall and CRC-specific mortality using Cox proportional hazards models, with adjustments for mortality-associated risk factors and other plasma markers of inflammation. We also assessed the relationship between levels of MIC1 and levels of prostaglandin-endoperoxide synthase 2 expression (PTGS2 or cyclooxygenase-2), measured in 245 tumor samples by immunohistochemistry. Compared with participants in the lowest quartile for plasma level of MIC1, the multivariate hazard ratio for CRC-specific death for participants in the highest quartile of MIC1 level was 2.40 (95% confidence interval: 1.33-4.34; P for linear trend = .009). The association of MIC1 with survival varied with level of PTGS2 expression in tumor samples (Pinteraction = .04). For individuals with PTGS2-positive tumors, the hazard ratio for CRC-specific death among those with high levels of MIC1 (equal to or greater than the median) was 2.13 (95% confidence interval: 0.99-4.58) compared with participants with low levels of MIC1 (below the median). In individuals with PTGS2-negative CRC, a high level of MIC1 was not associated with an increased risk of CRC-specific death (multivariate hazard ratio = 0.61; 95% confidence interval: 0.13-2.93). Based on an analysis of blood and colorectal tumor samples from 2 large studies, high plasma levels of MIC1 (growth differentiation factor 15) before diagnosis of CRC are associated with greater CRC-specific mortality, particularly in individuals with PTGS2-positive tumors.

Macrophage inhibitory cytokine 1 biomarker serum immunoassay in combination with PSA is a more specific diagnostic tool for detection of prostate cancer.

BackgroundProstate cancer (PCa) is the most common malignancy among men in the United States. Though highly sensitive, the often-used prostate-specific antigen (PSA) test has low specificity which leads to overdiagnosis and overtreatment of PCa. This paper presents results of a retrospective study that indicates that testing for macrophage inhibitory cytokine 1 (MIC-1) concentration along with the PSA assay could provide much improved specificity to the assay.MethodsThe MIC-1 serum level was determined by a novel p-Chip-based immunoassay run on 70 retrospective samples. The assay was configured on p-Chips, small integrated circuits (IC) capable of storing in their electronic memories a serial number to identify the molecular probe immobilized on its surface. The distribution of MIC-1 and pre-determined PSA concentrations were displayed in a 2D plot and the predictive power of the dual MIC-1/PSA assay was analyzed.ResultsMIC-1 concentration in serum was elevated in PCa patients (1.44 ng/ml) compared to normal and biopsy-negative individuals (0.93 ng/ml and 0.88 ng/ml, respectively). In addition, the MIC-1 level was correlated with the progression of PCa. The area under the receiver operator curve (AUC-ROC) was 0.81 providing an assay sensitivity of 83.3% and specificity of 60.7% by using a cutoff of 0.494 for the logistic regression value of MIC-1 and PSA. Another approach, by defining high-frequency PCa zones in a two-dimensional plot, resulted in assay sensitivity of 78.6% and specificity of 89.3%.ConclusionsThe analysis based on correlation of MIC-1 and PSA concentrations in serum with the patient PCa status improved the specificity of PCa diagnosis without compromising the high sensitivity of the PSA test alone and has potential for PCa prognosis for patient therapy strategies.