Tissue Tumor Necrosis Research Articles

Nicotinamide mononucleotide induces autophagy and ferroptosis via AMPK/mTOR pathway in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a common malignancy worldwide. Herein, we investigated the role of nicotinamide mononucleotide (NMN) in HCC progression. HCC cells were treated with NMN (125, 250, and 500 μM), and then nicotinamide adenine dinucleotide (NAD+ ) and NADH levels in HCC cells were measured to calculate NAD+ /NADH ratio. Cell proliferation, apoptosis, autophagy and ferroptosis were determined. AMPK was knocked down to confirm the involvement of AMPK/mTOR signaling. Furthermore, tumor-inhibitory effect of NMN was investigated in xenograft models. Exposure to NMN dose-dependently increased NAD+ level and NAD+ /NADH ratio in HCC cells. After NMN treatment, cell proliferation was inhibited, whereas apoptosis was enhanced in both cell lines. Additionally, NMN dose-dependently enhanced autophagy/ferroptosis and activated AMPK/mTOR pathway in HCC cells. AMPK knockdown partially rescued the effects of NMN in vitro. Furthermore, NMN treatment restrained tumor growth in nude mice, activated autophagy/ferroptosis, and promoted apoptosis and necrosis in tumor tissues. The results indicate that NMN inhibits HCC progression by inducing autophagy and ferroptosis via AMPK/mTOR signaling. NMN may serve as a promising agent for HCC treatment.

A holistic physics-informed neural network solution for precise destruction of breast tumors using focused ultrasound on a realistic breast model.

This study presented a novel approach for the precise ablation of breast tumors using focused ultrasound (FUS), leveraging a physics-informed neural network (PINN) integrated with a realistic breast model. FUS has shown significant promise in treating breast tumors by effectively targeting and ablating cancerous tissue. This technique employs concentrated ultrasonic waves to generate intense heat, effectively destroying cancerous tissue. In previous finite element method (FEM) models, the computational demands of handling extensive datasets, multiple dimensions, and discretization posed significant challenges. Our PINN-based solution operated efficiently in a mesh-free domain, achieving remarkable accuracy with significantly reduced computational demands, compared to conventional FEM techniques. Additionally, employing PINN for estimating partial differential equations (PDE) solutions can notably decrease the enormous number of discretized elements needed. The model employed a bowl-shaped acoustic transducer to focus ultrasound waves accurately on the tumor location. The simulation results offered detailed insights into each step of the FUS treatment process, including the generation of acoustic waves, the targeting of the tumor, and the subsequent heating and ablation of cancerous tissue. By applying a 3.8 nm displacement amplitude of transducer input pulse at a frequency of 1.1 MHz for 1 second, the temperature at the focal point elevated to 38.4 ℃, followed by another 90 seconds of cooling time, which resulted in significant necrosis of the tumor tissues. Validation of the PINN model's accuracy was conducted through FEM analysis, aligning closely with real-world FUS therapy scenarios. This innovative model provided physicians with a predictive tool to estimate the necrosis of tumor tissue, facilitating the customization of FUS treatment strategies for individual breast cancer patients.

Endoplasmic reticulum-mitochondrial calcium transport contributes to soft extracellular matrix-triggered mitochondrial dynamics and mitophagy in breast carcinoma cells.

Although mitochondrial morphology and function are considered to be closely related to matrix stiffness-driven tumor progression, it remains poorly understood how extracellular matrix (ECM) stiffness affects mitochondrial dynamics and mitophagy. Here, we found that soft substrate triggered calcium transport by increasing endoplasmic reticulum (ER) calcium release and mitochondrial (MITO) calcium uptake. ER-MITO calcium transport promoted the recruitment of dynamin-related protein 1 (Drp1) to mitochondria and phosphorylation at the serine 616 site, which induced mitochondrial fragmentation and Parkin/PINK1-mediated mitophagy. Furthermore, in vivo experiments demonstrated that soft ECM enhanced calcium levels in tumor tissue, Drp1 activity was required for soft ECM-induced mitochondrial dynamics impairment, and inhibition of Drp1 activity enhanced soft ECM-induced tumor necrosis. In conclusion, we revealed a new mechanism whereby ER-MITO calcium transport regulated mitochondrial dynamics and mitophagy through Drp1 translocation in response to soft substrates. These findings provide valuable insights into ECM stiffness as a potential target for antitumor therapy. STATEMENT OF SIGNIFICANCE: : Here, we examined the relationship between substrate stiffness and mitochondrial dynamics by using polyacrylamide (PAA) substrates to simulate the stages of breast cancer or BAPN to reduce tumor tissue stiffness. The results elucidated that soft substrate triggered the recruitment of DRP1 and subsequent mitochondrial fission and mitophagy by ER-MITO calcium transport. Furthermore, mitophagy partly attenuated soft ECM-mediated tumor tissue necrosis and contributed to tumor survival in vivo. Our discoveries revealed the molecular mechanisms by which mechanical stimulation regulates mitochondrial dynamics, providing valuable insights into ECM stiffness as a target for anti-tumor approaches, which could be beneficial for both biomechanics research and clinical applications.

Renal arterial infusion of tempol prevents medullary hypoperfusion, hypoxia, and acute kidney injury in ovine Gram-negative sepsis.

Renal medullary hypoperfusion and hypoxia precede acute kidney injury (AKI) in ovine sepsis. Oxidative/nitrosative stress, inflammation, and impaired nitric oxide generation may contribute to such pathophysiology. We tested whether the antioxidant and anti-inflammatory drug, tempol, may modify these responses. Following unilateral nephrectomy, we inserted renal arterial catheters and laser-Doppler/oxygen-sensing probes in the renal cortex and medulla. Noanesthetized sheep were administered intravenous (IV) Escherichia coli and, at sepsis onset, IV tempol (IVT; 30 mg kg-1 h-1 ), renal arterial tempol (RAT; 3 mg kg-1 h-1 ), or vehicle. Septic sheep receiving vehicle developed renal medullary hypoperfusion (76 ± 16% decrease in perfusion), hypoxia (70 ± 13% decrease in oxygenation), and AKI (87 ± 8% decrease in creatinine clearance) with similar changes during IVT. However, RAT preserved medullary perfusion (1072 ± 307 to 1005 ± 271 units), oxygenation (46 ± 8 to 43 ± 6 mmHg), and creatinine clearance (61 ± 10 to 66 ± 20 mL min-1 ). Plasma, renal medullary, and cortical tissue malonaldehyde and medullary 3-nitrotyrosine decreased significantly with sepsis but were unaffected by IVT or RAT. Consistent with decreased oxidative/nitrosative stress markers, cortical and medullary nuclear factor-erythroid-related factor-2 increased significantly and were unaffected by IVT or RAT. However, RAT prevented sepsis-induced overexpression of cortical tissue tumor necrosis factor alpha (TNF-α; 51 ± 16% decrease; p = 0.003) and medullary Thr-495 phosphorylation of endothelial nitric oxide synthase (eNOS; 63 ± 18% decrease; p = 0.015). In ovine Gram-negative sepsis, renal arterial infusion of tempol prevented renal medullary hypoperfusion and hypoxia and AKI and decreased TNF-α expression and uncoupling of eNOS. However, it did not affect markers of oxidative/nitrosative stress, which were significantly decreased by Gram-negative sepsis.

Delivery of sPD1 gene by anti-CD133 antibody conjugated microbubbles combined with ultrasound for the treatment of cervical cancer in mice

To explore new therapeutic options for cervical cancer, the inhibitory effect on cervical cancer of targeted CD133-loaded sPD1 gene microbubbles (MBs) combined with low-frequency ultrasound was studied and its mechanism was explored. We prepared microbubbles conjugated with anti-CD133 antibody to deliver the sPD1 gene and determined concentration, particle size, and potentials of MBs. In addition, we verified that CD133 targeted-MBs could specifically bind to U14 cervical cancer cells in vitro. A mouse model of subcutaneous xenograft cervical cancer was established and mice were divided into a control group, an non-targeted microbubble group, a CD133-MBs group, an sPD1-MBs group and a CD133/sPD1-MBs group. Compared with the control group, tumor growth was inhibited in each group, with the CD133/sPD1 group showing the strongest inhibitory effect after treatment. The tumor volume and weight inhibition rates in the CD133/sPD1-MBs group were 78.01% and 72.25% respectively, which were statistically different from the other groups (P < 0.05), and HE staining and TUNEL immunofluorescence showed necrosis and apoptosis in tumor tissue. Flow cytometry, lactate dehydrogenase, and indirect immunofluorescence experiments showed that T lymphocytes were activated and a large number of CD8-positive T cells infiltrated the tumor tissue after treatment, with the CD133/sPD1-MBs group showing the most prominent effects (P < 0.05). The combination of ultrasound with anti- CD133 antibody-conjugated microbubbles loaded with the sPD1 gene can inhibit the growth of cervical cancer, suggesting that the immunosuppressive microenvironment of the tumor is improved after treatment.

The positive effects of dietary arginine on juvenile hybrid snakehead (Channa maculata♀×Channa argus♂) fed high-carbohydrate diets: Liver inflammation antioxidant response, and glucose metabolism

The aim of this study was to investigate the effects of arginine supplementation on liver morphology, antioxidant capacity, inflammatory response, insulin signaling pathway and glucose metabolism in hybrid snakehead. Five experimental diets (2.31 %, 2.42 %, 2.72 %, 2.92 %, and 3.12 % arginine) were prepared by adding different levels of arginine to the basal diet containing high (27 %) level of carbohydrate. The results showed that the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and glutathione peroxidase (GSH-Px) of fish fed with arginine diets were significantly higher than those in control group (p < 0.05), accompanied by the lower malondialdehyde (MDA) content in the same groups (p < 0.05). There were obvious mild inflammatory cell infiltration and increased perihepatic pancreatic space in control group, while the pathological state of the liver was relieved in fish fed with arginine diets. Dietary arginine supplementation resulted in significantly higher glucose metabolism-related genes glucokinase (GK), phosphoenolpyruvate carboxkinase (PEPCK), glycogen synthase (GS) and pyruvate kinase (PK) and insulin signaling pathway key genes insulin receptor subset-1 (IRS-1), protein kinase B (Akt) and phosphoenolpyruvate carboxykinase (PI3K)(p < 0.05). Immune-related genes transforming growth factor β (TGF-β), heat shock protein 70 (HSP70) and heat shock protein 90 (HSP90) were significantly higher in the arginine supplemented diet than in the control group (p < 0.05). Immune-related genes interleukin-8 (IL8), interleukin-1β (IL1β) and tissue tumor necrosis factor-α (TNF-α) in fish fed with arginine diets were significantly lower than those in control group (p < 0.05). In conclusion, dietary arginine can alleviate the adverse effects of high carbohydrate diet on hybrid snakeheads by improving glucose metabolism and enhancing antioxidant, anti-inflammatory and non-specific immune functions.

Staphylococcal enterotoxin B as DNA vaccine against breast cancer in a murine model.

Recently, many efforts have been made to treat cancer using recombinant bacterial toxins and this strategy has been used in clinical trials of various cancers. Therapeutic DNA cancer vaccines are now considered as a promising strategy to activate the immune system against cancer. Cancer vaccines could induce specific and long-lasting immune responses against tumors. This study aimed to evaluate the antitumor potency of the SEB DNA vaccine as a new antitumor candidate against breast tumors in vivo. To determine the effect of the SEB construct on inhibiting tumor cell growth in vivo, the synthetic SEB gene, subsequent codon optimization, and embedding the cleavage sites were sub-cloned to an expression vector. Then, SEB construct, SEB, and PBS were injected into the mice. After being vaccinated, 4T1 cancer cells were injected subcutaneously into the right flank of mice. Then, the cytokine levels of IL-4 and IFN-γ were estimated by the ELISA method to evaluate the antitumor activity. The spleen lymphocyte proliferation, tumor size, and survival time were assessed. The concentration of IFN-γ in the SEB-Vac group showed a significant increase compared to other groups. The production of IL-4 in the group that received the DNA vaccine did not change significantly compared to the control group. The lymphocyte proliferation increased significantly in the mice group that received SEB construct than PBS control group (p < 0.001). While there was a meaningful decrease in tumor size (p < 0.001), a significant increase in tumor tissue necrosis (p < 0.01) and also in survival time of the animal model receiving the recombinant construct was observed. The designed SEB gene construct can be a new model vaccine for breast cancer because it effectively induces necrosis and produces specific immune responses. This structure does not hurt normal cells and is a safer treatment than chemotherapy and radiation therapy. Its slow and long-term release gently stimulates the immune system and cellular memory. It could be applied as a new model for inducing apoptosis and antitumor immunity to treat cancer.

Progress in the cryoablation and cryoimmunotherapy for tumor.

With the rapid advancement of imaging equipment and minimally invasive technology, cryoablation technology is being used more frequently in minimally invasive treatment of tumors, primarily for patients with early tumors who voluntarily consent to ablation as well as those with advanced tumors that cannot be surgically removed or cannot be tolerated. Cryoablation is more effective and secure for target lesions than other thermal ablation methods like microwave and radiofrequency ablation (RFA). The study also discovered that cryoablation, in addition to causing tumor tissue necrosis and apoptosis, can facilitate the release of tumor-derived autoantigens into the bloodstream and activate the host immune system to elicit beneficial anti-tumor immunological responses against primary. This may result in regression of the primary tumor and distant metastasis. The additional effect called " Accompanying effects ". It is the basis of combined ablation and immunotherapy for tumor. At present, there is a lot of research on the mechanism of immune response induced by cryoablation. Trying to solve the question: how positively induce immune response. In this review, we focus on: 1. the immune effects induced by cryoablation. 2. the effect and mechanism of tumor immunotherapy combined with cryoablation. 3.The clinical research of this combination therapy in the treatment of tumors.

Molecular cloning, expression analysis and immune-related functional identification of tumor necrosis factor alpha (TNFα) in Sepiella japonica under bacteria stress

Tumor necrosis factor α (TNFα), a cytokine mainly secreted by active macrophages and monocytes, causes hemorrhagic necrosis of tumor tissues, kills tumor cells, regulates inflammatory responses, and plays a crucial role in innate immunity. In this study, TNFα of Sepiella japonica (named as SjTNFα) was acquired, whose full-length cDNA was 1206 bp (GenBank accession no. ON357428), containing a 5′ UTR of 185 bp, a 3′ UTR of 137 bp and an open reading frame (ORF) of 1002bp to encode a putative peptide of 333 amino acids for constructing the transmembrane domain and the cytoplasmic TNF domain. Its predicted pI was 8.69 and the theoretical molecular weight was 44.72 KDa. Multiple sequence alignment and phylogenetic analysis showed that SjTNFα had the highest homology to Octopus sinensis, they fell into a unified branch and further clustered with other animals. Real-time PCR indicated that SjTNFα was widely expressed in all subject tissues, including spleen, pancreas, gill, heart, brain, optic lobe, liver and intestine, and exhibited the highest in the liver and the lowest in the brain. The relative expression of SjTNFα varied at the developmental period of juvenile stage, pre-spawning and oviposition in the squid, with the highest in the liver at the juvenile stage and oviposition, and in the optic lobe of pre-spawning. After being infected with Vibrio parahaemolyticus and Aeromonas hydrophila, the expression of SjTNFα in liver and gill were both upregulated with time, and the highest expression appeared at 24 h and 8 h in liver for different infection, and at 4 h in gill consistently. Cell localization showed that SjTNFα distributed on membrane of HEK293 cells because it was a type II soluble transmembrane protein. When HEK293 cells were stimulated with LPS of different concentrations, the NF-κB pathway was activated in the nucleus and the corresponding mRNA was transferred through the intracellular signal transduction pathway, resulting in the synthesis and release of TNFα, which made the expression of SjTNFα was up-regulated obviously. These findings showed that SjTNFα might play an essential role in the defense of S. japonica against bacteria challenge, which contributed to the understanding of the intrinsic immune signaling pathway of Cephalopoda and the further study of host-pathogen interactions.

Protective effect of rosmarinic acid-rich extract from Trichodesma khasianum Clarke against microbiota dysbiosis in high-fat diet-fed obese mice

Hypertrophy of adipose tissues and dysbiosis are hallmarks of obesity. Although drugs are applied for obesity treatment, side effects limit their use. The anti-obesity capacity of rosmarinic acid (RA) has been documented. Trichodesma khasianum Clarke is an edible RA-rich plant grown in Taiwan. Our previous study found that an 80 % ethanol extract of T. khasianum Clarke leaves (80EETC) ameliorates gastric mucosal damage through its anti-inflammatory, antioxidant, and microbiota modulation abilities. However, the anti-obesity effect of 80EETC remains unclear. Therefore, the objective of this study was to explore the protective effects of low-dose 80EETC (125 mg/kg b.w., 80EETCL) or high-dose 80EETC (250 mg/kg b.w., 80EETCH) on obesity development through gut microbiota modulation in high-fat diet (HFD)-induced C57BL/6 mice. The results showed a high RA content (89.2 ± 7.4 mg/g) in 80EETC. 80EETC administration significantly decreased body weight, body fat ratio, serum lipid levels (TC, TG, and LDL-C), adipose tissue accumulation, malondialdehyde (MDA), and tumor necrosis factor-α (TNF-α) in HFD-fed mice. Furthermore, supplementation with 80EETC reduced the Firmicutes/Bacteroidetes ratio and enhanced the relative abundance of gut microbiota (p_Bacteroidetes, f_Lactobacillus, f_Muribaculaceae, f_Prevotellaceae, g_Lactobacillus, g_Prevotellaceae_NK3B31_group, g_Ruminococcaceae_UCG–013, and g_Ruminococcaceae_UCG–014), which negatively correlated with obesity-related factors such as body weight, energy intake, fat accumulation in adipose tissue, TC, TG, LDL, and MDA. In conclusion, RA-rich 80EETC had a protective effect against obesity development and it has potential in healthy food applications.

ET-6 EFFICACY OF NEAR-INFRARED PHOTOIMMUNOTHERAPY TARGETING PODOPLANIN IN MALIGNANT GLIOMAS

Abstract Background Near-infrared photoimmunotherapy (NIR-PIT) is a new cancer treatment based on a conjugate of photosensitizer IR700 and an antibody which selectively binds to the surface antigen of cancer cells. Upon exposure to near infrared light, which does not harm normal tissues, cancer cells are selectively killed. Clinical application for EGFR amplified head and neck cancer has already been realized but is still in the preclinical stages of research for brain tumors. Finding a surface antigen which is selective to cancer cells is key to the effective application of NIR-PIT. In IDH-wildtype gliomas, we found that podoplanin (PDPN) is a promising candidate. PDPN is a transmembrane sialoglycoprotein highly expressed in IDH-wildtype gliomas and other cancers. Methods Three PDPN-positive (NGT-11, NGT-41, LN319) and three PDPN-negative (T98G, U87MG, U251MG) glioblastoma cell lines were treated with PDPN-IR700 conjugate and exposed to near infrared light. Cytotoxicity was assessed by cell viability assays. Next, we treated subcutaneous mouse xenografts by NIR-PIT. Results Cytotoxicity was only observed after NIR-PIT using PDPN-conjugate in cell lines expressing PDPN. This cytotoxicity was dose-dependent to near infrared light exposure, and most pronounced in LN319 cell line, which expressed the highest levels of PDPN. Time lapse videos revealed instantaneous expanding, followed by rounding of dead cells after treatment. NIR-PIT treated subcutaneous xenografts showed significantly smaller tumor sizes compared to control, and pathological investigation revealed selective and extensive necrosis of tumor tissue. Discussion/conclusion PDPN is highly expressed in IDH-wildtype gliomas, and we found that NIR-PIT was highly effective in treating PDPN expressing glioblastoma cell lines. NIR-PIT is anticipated to be more selective and cytotoxic than photodynamic therapy and is a promising new treatment strategy for IDH-wildtype gliomas.

Neuroprotective Effects of Milrinone on Acute Traumatic Brain Injury

Traumatic brain injury is still an important health problem worldwide. Traumatic brain injury not only causes direct mechanical damage to the brain but also induces biochemical changes that lead to secondary nerve cell loss. In this study, we investigated the neuroprotective effect of milrinone after traumatic brain injury (TBI) in a rat model. Forty male Wistar albino rats, were used. Rats were divided into 4 groups: 1) sham, 2) TBI, 3) TBI+Ringers, and 4) TBI+Milrinone. In group 1 (sham), only craniotomy was performed. In group 2 (TBI), TBI was performed after craniotomy. In group 3 (TBI+Ringer), TBI was performed after craniotomy and intraperitoneal Ringers solution was given immediately afterward. Group 4 (TBI+Milrinone), TBI was performed after craniotomy, and milrinone was given 1.0mg/kg milrinone intraperitoneally directly (0.5mg/kg milrinone intraperitoneally again 24hours, 48hours, and 72hours after trauma). Tests were performed for neurological and neurobehavioral functions. Immunohistochemistry and histopathology studies were performed. In group 4 compared with group 2 and group 3 groups, tests for neurological functions and neurobehavioral functions were significantly better. In the milrinone treatment used in group 4, plasma and brain tissue tumor necrosis factor, 8-OH 2-deoxyguanosine , and interleukin 6 levels were significantly decreased, and increased plasma and tissue IL-10 levels were detected. Histopathological spinal cord injury and apoptotic index increased in groups 2 and 3, while significantly decreasing in group4. This study shows for the first time that the anti-inflammatory, antioxidant and antiapoptotic properties of milrinone may be neuroprotective after TBI.

18F-5-FPN: A Specific Probe for Monitoring Photothermal Therapy Response in Malignant Melanoma.

Previously, we successfully synthesized a 18F-labeled positron-emission tomography (PET) tracer, termed 18F-5-fluoro-N-(2-[diethylamino]ethyl)picolinamide (18F-5-FPN), with high specificity for melanin. In this study, we sought to investigate the value of 18F-5-FPN in assessing the response to photothermal therapy (PTT) in melanoma via comparison with 18F-fluorodeoxyglucose (18F-FDG) to reveal an early response, recognize early recurrence, and distinguish the inflammatory response during the treatment. B16F10, inflammatory, and MDA-MB-231 models were subjected to 18F-FDG PET and 18F-5-FPN PET static acquisitions. We compared quantitative data to assess the specificity of different agents for different diseases. B16F10 and MDA-MB-231subcutaneous tumor models were irradiated with an 808 nm laser for PTT. Their survival was documented to observe the efficacy of and response to PTT, using 18F-5-FPN and 18F-FDG PET. 18F-5-FPN accumulated in B16F10 cell xenografts only, whereas 18F-FDG accumulated in all three models. Melanin in B16F10 cell xenografts successfully transformed the optical energy into heat. Hematoxylin and eosin (H&E) staining at 24 h revealed destruction and extensive necrosis of tumor tissue. PTT rapidly inhibited the growth of B16F10 cell xenografts and prolonged the median survival. The mean tumor uptakes of 18F-5-FPN on day 2 (7.52 ± 3.65 %ID/g) and day 6 (10.22 ± 6.00 %ID/g) were much lower than that before treatment (18.33 ± 4.98 %ID/g, p < 0.01). However, a significant difference in 18F-FDG uptakes was not found between day 1 after PTT and before treatment. Compared with 18F-FDG, 18F-5-FPN PET could estimate PTT efficacy in melanoma, monitor minimal recurrence, and distinguish melanoma from inflammation and other carcinoma types, thanks to its high affinity to melanin. 18F-5-FPN may provide a new approach for precise and accurate evaluation of response, timely management of therapeutic regimens, and sensitive follow-up.

OR04-6 Comprehensive Profiling of the MELK Inhibitor OTSSP167's Action and Its Effect on Sensitization to Wee1 Inhibitor AZD1775 in a Spectrum of ACC Models

Abstract Adrenocortical carcinoma (ACC) is an aggressive cancer with a significant risk of recurrence and high mortality rates. ACC has limited response to currently used treatments and there is an unmet clinical need to identify new treatment options. Disease rarity coupled with absence of preclinical models of disease heterogeneity contributed to limited therapeutic progress in ACC. We previously established new in vitro and in vivo ACC models (CUACC1, CUACC2, CUACC9) with variable genomic alterations commonly seen in ACC tumors. Prior work demonstrated that maternal leucine embryonic kinase (MELK) is an oncogenic kinase in ACC, and that it can be targeted with the MELK inhibitor OTSSP167. To further evaluate the anti-tumor efficacy and mode of action of OTSSP167, this study used genetically diverse ACC cell lines (CUACC1, CUACC2, H295R), patient derived xenografts (PDX) (CUACC1 and CUACC9) and a cell line xenograft model (H295R) of ACC. ACC PDXs treated with 10 mg/kg OTSSP167 had significantly reduced MELK expression, tumor growth inhibition of more than 80%, reduced mitosis, and increased tumor tissue necrosis. Transcriptomic and phospho-proteomic analysis at multiple treatment timepoints in in vitro models revealed that OTSSP167 induced wide transcriptomic and proteomic changes involving effectors of signal transduction, cell cycle, protein translation, and epigenetic regulation to exert its anti-tumorigenic effects. Evaluation of OTSSP167's role in protein synthesis via a puromycin based SUnSET (surface sensing of translation) and kinase assay established that OTSSP167 inhibits de-novo protein synthesis, and targets RSK1 and its substrate rpS6 (S235/S236). Varying the degree of MELK silencing in H295R cell lines using a doxycycline inducible shRNA construct, inhibited RSK1 phosphorylation at T573 and S380 indicating a MELK-RSK1 axis in ACC. Additionally, OTSSP167's effect on the DNA damage pathway was validated, which activated the G2/M checkpoint protein Wee1, increased inhibitory phosphorylation of CDK1, causing an accumulation of cyclin B1 and G2/M arrest independent of TP53 status. Combination treatment with the Wee1 inhibitor AZD1775 and OTSSP167 enhanced OTSSP167 mediated cytotoxic effects in in-vitro models with an increase in mitotic entry and caspase mediated apoptosis. In summary, OTSSP167 exerted its anti-tumorigenic effects via inhibition of the protein translation machinery, DNA damage and cell cycle dysregulation causing drug sensitization. Results from these studies are a first demonstration of the anti-cancer effects of OTSSP167 in sensitization to Wee1 inhibition in ACC. These data suggest that OTSSP167 is a potential therapeutic option for patients with ACC and could be combined with Wee1 inhibitors in a clinical trial using a synthetic lethal approach. Presentation: Saturday, June 11, 2022 12:45 p.m. - 1:00 p.m.

Trastuzumab-based near-infrared photoimmunotherapy in xenograft mouse of breast cancer.

Near-infrared photoimmunotherapy (NIR-PIT) is a novel form of cancer treatment using conjugates of antibody against overexpressed antigens in cancers and photoabsorber IRDye700DX. HER2 is overexpressed in various cancers, for which molecular targeted therapy such as trastuzumab has been developed. The present study investigated the efficacy potential of HER2-targeted NIR-PIT using trastuzumab-IRDye700DX conjugate (Tra-IR700) in HER2-positive breast cancer. We first examined the reactivity of Tra-IR700 and the cytotoxicity of NIR-PIT in vitro. HER2-positive BT-474 and SK-BR-3 cells and HER2-negative BT-20 cells were used. Tra-IR700 fluorescence was only observed in HER2-positive breast cancer cell lines, and the fluorescence was localized to the cell surface. Furthermore, HER2-positive breast cancer cell lines treated with NIR-PIT showed swelling and blebbing shortly after irradiation, and eventually increased PI-positive dead cells. Next, tumor accumulation of Tra-IR700 and tumor damage by NIR-PIT were examined in vivo. Tra-IR700 was administered intravenously to a xenograft model in which BT-474 cells were implanted subcutaneously in BALB/c nude mice. Tra-IR700 fluorescence was the highest in tumor tissue 1day after administration, and the fluorescence was localized to the cell membrane of tumor cells. At this time point, NIR-PIT resulted in diffuse necrosis of tumor tissues 1day after irradiation. These results suggest that NIR-PIT with Tra-IR700 induces a highly selective therapeutic effect in a HER2-positive breast cancer model. NIR-PIT using Tra-IR700 is expected to be a novel treatment for HER2-positive cancers, including breast cancer.

Fenofibrate ameliorates letrozole-induced polycystic ovary in rats via modulation of PPARα and TNFα/CD95 pathway.

Polycystic ovary syndrome (PCOS) is a prevalent endocrine health problem during the childbearing period that seriously affects fertility in females. Fenofibrate, a peroxisome proliferator-activated receptor-alpha (PPARα) agonist, showed beneficial effects in models of endocrine disturbances. Thus, we evaluated the potential therapeutic effect of fenofibrate in experimental PCOS. Rats received oral fenofibrate (300 mg/kg/day) for three weeks following a three-week PCOS induction regimen using oral letrozole (1 mg/kg/day). We determined the changes in body weight, levels of serum testosterone, insulin, anti-Müllerian hormone (AMH), ovarian malondialdehyde (MDA), superoxide dismutase (SOD), and tissue tumor necrosis factor-alpha (TNFα) and CD95 protein expressions. The tissue expression of interleukin-10 (IL10) and PPARα genes was determined. Letrozole-treated rats showed successful induction of PCOS, confirmed by histopathology and significantly increased body weight, testosterone, insulin, AMH, and MDA, and decreased SOD. Ovaries of untreated PCOS rats showed increased TNFα and CD95 and decreased PPARα and IL10 expression. Administration of fenofibrate ameliorated the letrozole-induced PCOS changes. Fenofibrate-mediated amelioration of PCOS in rats is attributed partly to its antioxidant, anti-inflammatory, and anti-apoptotic properties and activation of PPARα.

Numerical study of magnetic hyperthermia ablation of breast tumor on an anatomically realistic breast phantom.

Magnetic fluid hyperthermia (MFH) is a novel reliable technique with excellent potential for thermal therapies and treating breast tumours. This method involves injecting a magnetic nanofluid into the tumour and applying an external AC magnetic field to induce heat in the magnetic nanoparticles (MNPs) and raise the tumour temperature to ablation temperature ranges. Because of the complexity of considering and coupling all different physics involves in this phenomenon and also due to the intricacy of a thorough FEM numerical study, few FEM-based studies address the entire MFH process as similar to reality as possible. The current study investigates a FEM-based three-dimensional numerical simulation of MFH of breast tumours as a multi-physics problem. An anatomically realistic breast phantom (ARBP) is considered, some magnetic nanofluid is injected inside the tumour, and the diffusion phenomenon is simulated. Then, the amount of heat generated in the MNP-saturated tumour area due to an external AC magnetic field is simulated. In the end, the fraction of tumour tissue necrotized by this temperature rise is evaluated. The study’s results demonstrate that by injecting nanofluid and utilizing seven circular copper windings with each coil carrying 400 A current with a frequency of 400 kHz for generating the external AC magnetic field, the temperature in tumour tissue can be raised to a maximum of about 51.4°C, which leads to necrosis of entire tumour tissue after 30 minutes of electromagnetic field (EMF) exposure. This numerical platform can depict all four various physics involved in the MFH of breast tumours by numerically solving all different equation sets coupled together with high precision. Thus, the proposed model can be utilized by clinicians as a reliable tool for predicting and identifying the approximate amount of temperature rise and the necrotic fraction of breast tumour, which can be very useful to opt for the best MFH therapeutic procedure and conditions based on various patients. In future works, this numerical platform’s results should be compared with experimental in-vivo results to improve and modify this platform in order to be ready for clinical applications.

P11.68.A Hemangiopericytoma, case series and clinic-pathological analysis

Abstract Background Hemangiopericytoma (HPC) is an extremely rare aggressive tumor of mesenchymal origin, which constitutes less than 1% of primary tumors of the central nervous system. Typically it occurs in young adults (mean age at diagnosis 30-50 years). Initially described as gemmangiomas, was first called hemangiopericytoma in 1942 but it was considered as a variant of meningioma until 1993 when it was finally recognized as a distinct clinical-pathological entity by the World Health Organization (WHO). In 2016, there was the unification of both HPC and Solitary Fibrous Tumor (SFT) in a single spectrum of rare and thick mesenchymal tumors due to the identification, in both tumoral types, of a molecular alteration called NAB2-STAT6 fusion. Material and Methods The present study is a case series of patients that received a pathological diagnosis of SFT/HPC at our Institute from 1981 to 2020. Clinical data included: demographic data, tumor characteristics and treatment modality. Hystological diagnosis and immunohistochemistry allowed to confirm diagnosis, based on the 2016 WHO classification. Haematoxylin staining was completed with immunohistochemistry with primary antibodies for STAT 6 and evaluation of presence of tumor necrosis. Statistical analysis was conducted considering age at first diagnosis, treatment modality, extent of tumor resection, presence of necrosis in histological analysis compared to PFS-1, OS and relapses number. Results The case series was composed by 82 patients (46 men and 36 women) with median age of 43 (± 20) that accede to our Institute from 1981 to 2020. Our statistical analysis showed significative correlation between age, extend of tumor resection and presence of necrosis with OS and relapses number. Conclusion Hemangiopericytoma is a very rare cerebral tumors, and for this reason large case series are not disposable in literature. In our study we collected all disposable data of every patient with pathological diagnosis of SFT/HPC acceded to our hospital in the long period of 39 years in the aim to verify the presence of significative correlations. We observed strong correlation between tumor resection and presence of necrosis in tumor tissue with prognosis parameters (OS and number of relapses). This result it could be important to conduct the decisional process about treatment after surgical resection, considering also pathological features, rather than only surgical resection.

Influence of Maternal Ethanol Exposure on Systemic Hemodynamic Variables and Histopathological Changes in the Aorta Wall of Male Rat Offspring: A Three-month Follow-up.

Background: Alcohol consumption in pregnancy is associated with an increased risk of cardiovascular abnormalities, but the mechanisms are unknown. This study evaluated the impact of ethanol exposure on the offspring’s aorta structural, functional, and molecular alterations on postnatal (PN) both on days 21 and 90.Methods:This experimental study was conducted at Urmia University of Medical Sciences (Urmia, Iran) in 2019. Twenty Pregnant Wistar rats on the seventh day of Gestation Day (GD) were randomly divided into two groups: control and ethanol-treated groups (n=10 per group). From the seventh day of GD throughout lactation, rats in the ethanol group were fed binge alcohol (4.5 g/Kg body weight) once daily. Systemic hemodynamic variables in the offspring were analyzed using waveform contour analysis 90 days after birth. On postnatal days (PN) 21 and 90, aorta wall histological alterations and the level of inflammatory factors were assessed in the aorta of male offspring. The statistical differences were examined via an independent samples t test. P<0.05 was considered to be statistically significant. Results: The results revealed that offspring in the ethanol group had higher systolic, diastolic, mean arterial pressure, and dicrotic pressure than the control group (P<0.001). The level of aorta tissue tumor necrosis factor (TNF)-α, intercellular adhesion molecule (ICAM)-1, nuclear factor (NF)-κ, and endothelin-1 were significantly higher in the ethanol offspring group than in the control group (P<0.001). Histopathological changes such as total aorta thickness, tunica media, tunica adventitia, elastin fiber thickness, fiber interval, and elastin/media ratio significantly increased in the aorta of the offspring of the ethanol group compared to the control group 21 and 90 days after birth.Conclusion: Our findings suggest that prenatal and early postnatal ethanol exposure-induced cardiovascular abnormalities are, in part, due to predisposing the aorta to atherosclerosis, which was mediated through the aorta wall remodeling and inflammation process.

The Efficacy of Fosbretabulin Disodium Combined with Radiofrequency Ablation in Lung Cancer.

Radiofrequency ablation (RFA) is a technology that uses radiofrequency thermal effect to induce coagulation necrosis of tumor tissue under the guidance of imaging. However, distant metastasis of tumor cells caused by tumor angiogenesis can lead to incomplete tumor clearing. In this study, LLC1 cell line was used for the construction of subcutaneous xenografts. Either 10 mg/kg or 20 mg/kg Fosbretabulin disodium (FBTD) was intragastrically administered every 2 days for a week. RFA was performed at the end of medication. The proportion of T cells was examined by flow cytometry. Serum IgG and IgA levels of mice were examined by ELISA. Expression of certain genes was estimated by qRT-PCR assay. In this study, we demonstrated that FBTD was able to significantly enhance RFA-induced immune function in tumor-bearing mice by upregulating RFA-induced CD8+ killer T cells. Consistently, 10 mg/kg or 20 mg/kg FBTD therapy upregulated the percentage of IFNγ+ and TNFα+ CD8+ tumor infiltrating lymphocytes in tumor-bearing mice compared to the RFA alone or FBTD alone group. Mechanistically, we reported that FBTD inhibited the RFA-induced PD-1 and PD-L1 upregulation in vivo. In conclusion, we demonstrated that FBTD promoted the antitumor effects of RFA in lung tumor-bearing mice in this study.