Growth Of Solid Tumors Research Articles

An Acid-Responsive Iron-Based Nanocomposite for OSCC Treatment

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer, characterized by invasiveness, local lymph node metastasis, and poor prognosis. Traditional treatment and medications have limitations, making the specific inhibition of OSCC growth, invasion, and metastasis a challenge. The tumor microenvironment exhibits mildly acidity and high concentrations of H2O2, and its exploitation for cancer treatment has been widely researched across various cancers, but research in the oral cancer field is relatively limited. In this study, by loading ultra-small Prussian blue nanoparticles (USPBNPs) into mesoporous calcium–silicate nanoparticles (MCSNs), we developed an acid-responsive iron-based nanocomposite, USPBNPs@MCSNs (UPM), for the OSCC treatment. UPM demonstrated excellent dual enzyme activities, generating toxic ·OH in a mildly acidic environment, effectively killing OSCC cells and producing O2 in a neutral environment to alleviate tissue hypoxia. The results showed that UPM could effectively inhibit the proliferation, migration, and invasion of OSCC cells, as well as the growth of mice solid tumors, without obvious systemic toxicity. The mechanisms may involve UPM inducing ferroptosis of OSCC cells by downregulating the xCT/GPX4/glutathione (GSH) axis, characterized by intracellular iron accumulation, reactive oxygen species accumulation, GSH depletion, lipid peroxidation, and abnormal changes in mitochondrial morphology. Therefore, this study provides empirical support for ferroptosis as an emerging therapeutic target for OSCC and offers a valuable insight for future OSCC treatment.

Selenium suppressed growth of Ehrlich solid tumor and improved health of tumor-bearing mice.

Selenium (Se) is an important micronutritional biomolecule in cancer therapy. The current work evaluated the anticancer effect of Se and its ability to improve health of mice with solid Ehrlich carcinoma implanted subcutaneously. Four groups of five female BALB/c mice each were assembled. Ehrlich tumor cells were engrafted into two of them, either with or without Se therapy. The other groups served as control groups, either with or without Se treatment. Se treatment resulted in a notable decrease in both tumor volume and animal body mass in tumor-bearing mice. Treatment with Se markedly increased oxidative stress in tumor while ameliorating oxidative stress in sera of tumors-bearing mice. Similarly, treatment with Se resulted in downregulation of inflammatory cytokines (TNF-α and IL-6) while increasing IL-10 in serum of tumor-bearing mice. Conversely, selenium increased TNF- α and IL-6 and decreased IL-10 in tumor suggesting disruption of tumor immunity. The increased oxidative stress and inflammation in tumor tissue dysregulated cell cycle phases with increase apoptotic tumor cells population in G0/G1 phase. This is supported by the increased levels apoptotic regulating proteins (Bax and caspase-3 and P-53) while decreasing Bcl-2 in the tumor tissue. Treatment with Se also resulted in increased comet parameters indicating DNA damage of tumor cells. Histopathological examination revealed a significant decrease in a number of neoplastic cells within tumor of mice that treated with Se. In conclusion, these findings suggest that Se therapy significantly suppressed solid tumor proliferation and growth while mitigating the health status of tumor-bearing mice.

Abstract 2908: EDB+FN is an attractive therapeutic target in oncology: Insights from protein expression analysis of solid tumors

Abstract Stroma plays a major role in the initiation, growth, survival, and drug-resistance of solid tumors, yet few therapeutics specifically target tumor-associated stroma. Extra-domain-B fibronectin (EDB+FN) is an oncofetal splice variant of fibronectin upregulated in solid tumor stroma, which is associated with tumor growth, angiogenesis, and metastases. Here we developed an immunohistochemistry (IHC) assay to assess EDB+FN protein expression in the stroma of tumor and normal tissues to characterize the potential of EDB+FN as a therapeutic target for solid tumors with high unmet need. We validated a primary detection antibody and titrated assay conditions to achieve specificity and a broad dynamic range of EDB+FN expression levels across indications, including thyroid cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, soft tissue sarcoma, pancreatic cancer, non-small cell lung cancer, ovarian cancer, and renal cancer. Considering the stroma-specific expression pattern of EDB+FN, we developed a scoring algorithm weighting both the intensity and distribution of EDB+FN expression specifically in the stroma compartment and separately in tumor cells. Using this assay and scoring approach, we screened more than 200 tumor samples from 10 solid tumor indications and found EDB+FN to be highly differentially expressed in tumor-associated stroma with little to no expression on tumor cells and trace to undetectable levels in healthy tissue. EDB+FN was expressed across the indications evaluated with a wide range of expression both within and between indications. Stromal EDB+FN expression was not correlated with stromal density, which also varied widely within and across indications. In parallel, we developed an algorithm using digital pathology to approximate pathologist scoring of stromal density and EDB+FN expression, allowing streamlined screening of preclinical samples for in vivo studies and indication selection. The disease-specific expression pattern of EDB+FN and high incidence of expression across indications make EDB+FN an ideal target for high-unmet-need solid tumors. The specificity and dynamic range of our IHC assay for detection of EDB+FN protein expression position it as a robust potential biomarker assay for EDB+FN-targeting therapeutics. Citation Format: Sara Lewandowski, Liyang Diao, Alyssa Quigley, Marsha Crochiere, Jan Pinkas. EDB+FN is an attractive therapeutic target in oncology: Insights from protein expression analysis of solid tumors [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 2908.

Abstract 6718: Evaluating toxicity and efficacy of bispecific EGFRXCD3 T cell engager in solid tumor-bearing humanized animal model

Abstract Background: Cancer immunotherapy has currently become a highly active research area of oncology and a common treatment strategy for several cancer types. One prominent reason is that the utilized animal models could not accurately predict the of toxic response of human immune system to anti-cancer drug. For improving the safety of immune drug development, it is urgent to establish a predictive nonclinical animal platform that can accurately predict human immune response to the drugs and detect potential unwanted toxicity to human patient will be critical. We have demonstrated a disseminated tumor-bearing humanized mouse model that can evaluate the toxicity and efficacy of bispecific T cell engager (BiTE). In this study, we co-engrafted disseminated and solid tumor cells to the same mice and compared with mice engrafted with disseminated tumor or solid tumor. The purpose is to develop a novel tumor-bearing mouse model that can simultaneously evaluate the long-term efficacy against solid tumors in addition to cytokine level assessment. Method: Epidermal growth factor receptor (EGFR) expressing human triple negative breast cancer MDA-MB-231/Luciferase-2A-GFP (MDA) cell line was utilized to establish solid tumors in mice. The cells were implanted into PBMC humanized NSG-MHC class I/II double knock out (DKO) mice via orthotopic injection into mammary fat pad (MFP). Disseminated MDA cells were injected intravenously (IV) and tumor cells normally colonize in mouse lungs. Following EGFRxCD3 BiTE dosing initiation, tumor burden in MFP and lungs were evaluated via in vivo Xenogen imaging system, and solid tumor volume in MFP was measured 2 to 3 times a week by a digital caliper. To evaluate the toxicity in response to EGFRxCD3 BiTE treatment, human inflammatory cytokine levels was analyzed by BD cytometric bead. Clinical observations and body weight change, and human immune cell activation were evaluated. Result: EGFRxCD3 BiTE treatments significantly inhibited MFP solid tumor growth regardless the presence of IV implanted with MDA cells. EGFRxCD3 BiTE treatment induced elevated human inflammatory cytokines from humanized mice implanted with combined MFP and IV tumors. By comparison, the serum cytokine levels were low in mice implanted with MDA cells via MFP alone, suggesting that disseminated tumor cells are required for cytokine production induced by EGFRxCD3 BiTE. Conclusion: Without disseminated tumor, MFP solid tumor-bearing mice can be served as a model for drug efficacy but did not induce inflammatory cytokines by EGFRxCD3 BiTE treatment. Combination of solid and disseminated tumor models in the same mice provide a new approach for simultaneously evaluating the toxic cytokine release and efficacy of immunotherapy drugs. Citation Format: Guoxiang Yang, Li-Chin Yao, James G. Keck, Mingshan Cheng. Evaluating toxicity and efficacy of bispecific EGFRXCD3 T cell engager in solid tumor-bearing humanized animal model [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 6718.

Abstract 742: PYX-201, a stroma-targeting ADC composed of an anti-EDB+FN antibody conjugated to Auristatin0101, demonstrates strong anti-tumor efficacy across multiple human cancer indications in pre-clinical PDX tumor models

Abstract Stroma is crucial to support solid tumor growth, metastasis and resistance to treatment. While most antibody-drug conjugates (ADCs) directly bind to cancer cells, PYX-201, an investigational drug, is designed to target tumor stroma by binding to the extra-domain B splice variant of fibronectin (EDB+FN), a matrix protein abundantly expressed in the tumor microenvironment of many solid tumors with absent or low expression in normal adult tissues. In vitro studies demonstrated PYX-201 was strongly cytotoxic to EDB+FN positive Caki2 cells, but not to EDB+FN negative HT29 cells. In addition, PYX-201 was 30-times more potent than a non-targeted ADC control, demonstrating PYX-201 cytotoxic effects were EDB+FN dependent. Furthermore, when Caki2 and HT29-luciferase cancer cells were co-cultured, the luciferase activity was used to quantify the amount of EDB+FN negative HT29 cells only. Interestingly, PYX-201 was able to kill EDB+FN negative cancer cells in a co-culture assay as shown by a decrease in luciferase activity, demonstrating PYX-201 bystander activity. Pre-clinical mouse model studies were conducted to evaluate PYX-201 anti-tumor efficacy and pharmacokinetic (PK) properties. First, a comprehensive patient-derived xenograft (PDX) mouse model trial was designed to evaluate the anti-tumor efficacy of PYX-201. To fully represent the diversity and characteristics of human tumors, PDX models were pre-selected from ten solid tumor indications based on different levels of EDB+FN protein expression and stromal density. PYX-201 (Q4D × 4 at 3mg/kg) was well tolerated, and strong anti-tumor responses (% Tumor Growth Inhibition [TGI] >90%) were observed in PDX models across various cancer indications. Potential correlations between EDB+FN protein expression with anti-tumor activity will be evaluated. Remarkably, long-term anti-tumor efficacy of PYX-201 was observed as tumors did not relapse even after >100 days in a breast cancer PDX model. Additionally, plasma samples were analyzed from a cell line derived xenograft mouse model after a single dose of PYX-201 to determine its PK profile. At 3 mg/kg, the maximum concentration (Cmax) was 28.1 μg/mL, area under the curve (AUC) was 1,732.7 h*μg/mL, with a low clearance of 1.5 mL/h/kg, and the half-life of the mAb from PYX-201 was calculated at 81.2 h. These PK parameters were associated with strong PYX-201 anti-tumor efficacy (% TGI >90%) in the H1975 xenograft model. Altogether, PYX-201 was well tolerated and demonstrated strong anti-tumor efficacy in a variety of pre-clinical human PDX models. PYX-201 is a promising and innovative ADC which is currently under investigation in a Phase I clinical trial (NCT05720117). Citation Format: Nicolas Severe, Amanda Facklam, Liz McMichael, Biplab Das, Sara Lewandowski, Justin Trickett, Liyang Diao, Chengfeng Merriman, Chuan Shen, Jianwen Feng, Shawn Harriman, Marsha Crochiere, Philipp Steiner, Jan Pinkas. PYX-201, a stroma-targeting ADC composed of an anti-EDB+FN antibody conjugated to Auristatin0101, demonstrates strong anti-tumor efficacy across multiple human cancer indications in pre-clinical PDX tumor models [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 742.

Effects of Ethanol Concentrations on Primary Structural and Bioactive Characteristics of Dendrobium officinale Polysaccharides.

Ethanol fractional precipitation can initially separate polysaccharides according to the structure, which exhibits strong correlation with the biological activities. This study aimed to investigate the impact of varying ethanol concentrations on the structural characteristics, and the antitumor and antioxidant activities of polysaccharides derived from Dendrobium officinale through ethanol fractional precipitation, as well as their internal relationships. The polysaccharides acquired by absolute alcohol additions at a final liquor-ethanol volume ratio of 1:1, 1:2, and 1:4 were named DOP-1, DOP-2, and DOP-4, and the supernatant was named DOP-S. The results of the structural analysis revealed that the increase in ethanol concentrations resulted in a reduction in the molecular weights and the acetylation degree of the polysaccharides, as well as a decrease in mannose content and an increase in glucose content. In vitro experiments demonstrated that DOP-S exhibited optimal antitumor and antioxidant activities. Animal experiments further confirmed that DOP-S suppressed the growth of solid tumors significantly, enhanced lymphocytes, mediated immune ability, and improved the activity of antioxidant enzymes. These findings would establish a theoretical foundation and provide technical support for further advances and applications of polysaccharides derived from D. officinale in the fields of food and medicine.

Exploring Tumor-Promoting Qualities of Cancer-Associated Fibroblasts and Innovative Drug Discovery Strategies With Emphasis on Thymoquinone.

Tumor epithelial development and chemoresistance are highly promoted by the tumor microenvironment (TME), which is mostly made up of the cancer stroma. This is due to several causes. Cancer-associated fibroblasts (CAFs) stand out among them as being essential for the promotion of tumors. Understanding the fibroblastic population within a single tumor is made more challenging by the undeniable heterogeneity within it, even though particular stromal alterations are still up for debate. Numerous chemical signals released by tumors improve the connections between heterotypic fibroblasts and CAFs, promoting the spread of cancer. It becomes essential to have a thorough understanding of this complex microenvironment to effectively prevent solid tumor growth. Important new insights into the role of CAFs in the TME have been revealed by recent studies. The objective of this review is to carefully investigate the relationship between CAFs in tumors and plant secondary metabolites, with a focus on thymoquinone (TQ). The literature published between 2010 and 2023 was searched in PubMed and Google Scholar with keywords such as TQ, TME, cancer-associated fibroblasts, mechanism of action, and flavonoids. The results showed a wealth of data substantiating the activity of plant secondary metabolites, particularly TQ's involvement in blocking CAF operations. Scrutinized research also clarified the wider effect of flavonoids on pathways related to cancer. The present study highlights the complex dynamics of the TME and emphasizes the critical role of CAFs. It also examines the possible interventions provided by secondary metabolites found in plants, with TQ playing a vital role in regulating CAF function based on recent literature.

Solid tumor cohesion and growth are overcome in phagocytosis by SIRPα-knockout macrophages

Solid tumor cohesion and growth are overcome in phagocytosis by SIRPα-knockout macrophages

Inhibition of Bromodomain Proteins Enhances Oncolytic HAdVC5 Replication and Efficacy in Pancreatic Ductal Adenocarcinoma (PDAC) Models.

Pancreatic ductal adenocarcinoma (PDAC) is the most aggressive type of pancreatic cancer, which rapidly develops resistance to the current standard of care. Several oncolytic Human AdenoViruses (HAdVs) have been reported to re-sensitize drug-resistant cancer cells and in combination with chemotherapeutics attenuate solid tumour growth. Obstacles preventing greater clinical success are rapid hepatic elimination and limited viral replication and spread within the tumour microenvironment. We hypothesised that higher intratumoural levels of the virus could be achieved by altering cellular epigenetic regulation. Here we report on the screening of an enriched epigenetics small molecule library and validation of six compounds that increased viral gene expression and replication. The greatest effects were observed with three epigenetic inhibitors targeting bromodomain (BRD)-containing proteins. Specifically, BRD4 inhibitors enhanced the efficacy of Ad5 wild type, Ad∆∆, and Ad-3∆-A20T in 3-dimensional co-culture models of PDAC and in vivo xenografts. RNAseq analysis demonstrated that the inhibitors increased viral E1A expression, altered expression of cell cycle regulators and inflammatory factors, and attenuated expression levels of tumour cell oncogenes such as c-Myc and Myb. The data suggest that the tumour-selective Ad∆∆ and Ad-3∆-A20T combined with epigenetic inhibitors is a novel strategy for the treatment of PDAC by eliminating both cancer and associated stromal cells to pave the way for immune cell access even after systemic delivery of the virus.

Computational modeling of early-stage breast cancer progression using TPFA method: A numerical investigation

In this paper, a finite volume (TPFA) method is employed to simulate a degenerate breast cancer model that captures the progressive mutations from a normal breast stem cell to a tumor cell. The model incorporates a degenerate parabolic equation to represent the interaction between solid tumor growth and its environment, which involves the release of degradative enzymes governed by a partial differential equation. The discrete maximum principle is verified to be satisfied by the proposed finite volume scheme, and the convergence of the existing discrete solutions to a weak solution is proven. Additionally, the development of breast cancer is demonstrated through a numerical experiment.

P66ShcA promotes malignant breast cancer phenotypes by alleviating energetic and oxidative stress

Significant efforts have focused on identifying targetable genetic drivers that support the growth of solid tumors and/or increase metastatic ability. During tumor development and progression to metastatic disease, physiological and pharmacological selective pressures influence parallel adaptive strategies within cancer cell sub-populations. Such adaptations allow cancer cells to withstand these stressful microenvironments. This Darwinian model of stress adaptation often prevents durable clinical responses and influences the emergence of aggressive cancers with increased metastatic fitness. However, the mechanisms contributing to such adaptive stress responses are poorly understood. We now demonstrate that the p66ShcA redox protein, itself a ROS inducer, is essential for survival in response to physiological stressors, including anchorage independence and nutrient deprivation, in the context of poor outcome breast cancers. Mechanistically, we show that p66ShcA promotes both glucose and glutamine metabolic reprogramming in breast cancer cells, to increase their capacity to engage catabolic metabolism and support glutathione synthesis. In doing so, chronic p66ShcA exposure contributes to adaptive stress responses, providing breast cancer cells with sufficient ATP and redox balance needed to withstand such transient stressed states. Our studies demonstrate that p66ShcA functionally contributes to the maintenance of aggressive phenotypes and the emergence of metastatic disease by forcing breast tumors to adapt to chronic and moderately elevated levels of oxidative stress.

Metronomic cyclophosphamide and metformin inhibited tumor growth and repopulated tumor-infiltrating lymphocytes in an experimental carcinoma model

Metformin is a widely used antidiabetic biguanide. Retrospective data demonstrated the association of metformin use with survival benefit in multiple tumor types. Interest in repurposing metformin to treat cancer has not been translated into encouraging clinical benefit. In animal models, metformin activated cytotoxic T cells and exerted an immune-mediated anticancer effect. The current research was conducted to investigate the possible therapeutic benefit of metformin in combination with metronomic cyclophosphamide in an experimental cancer model. Ehrlich ascites carcinoma was injected into the subcutaneous tissue to induce solid tumors in syngeneic mice. Exponential solid tumor growth ensued and was effectively arrested with the administration of a cytotoxic dose of parenteral cyclophosphamide. Alternatively, oral metformin and continuous, low-dose cyclophosphamide significantly inhibited tumor growth relative to untreated mice. The drug combination was well tolerated. Histopathological examination of the tumor showed an increased number of tumor-infiltrating lymphocytes and enhanced expression of granzyme B by this drug combination. The current data suggests a potential role of metformin and metronomic chemotherapy that warrants further investigation.

The Human Placental Amniotic Membrane Mesenchymal-Stromal-Cell-Derived Conditioned Medium Inhibits Growth and Promotes Apoptosis of Human Cholangiocarcinoma Cells In Vitro and In Vivo by Suppressing IL-6/JAK2/STAT3 Signaling.

Mesenchymal stromal cells (MSCs) have recently been shown to play an important role in the growth and progression of many solid tumors, including cholangiocarcinoma (CCA). The human placental amniotic membrane (hPAM) is one of the most favorable sources of MSCs due to its availability and non-invasive harvesting procedure. However, the role of human placental amniotic membrane mesenchymal stromal cells (hPAMSCs) in the growth and progression of human CCA has not yet been determined. This study investigates the effects of conditioned medium derived from hPAMSCs (PA-CM) on the properties of three human CCA cell lines and explores possible mechanisms of action. Varying concentrations of PA-CM were used to treat CCA cells to determine their effects on the proliferation and apoptosis of CCA cells. The results showed that PA-CM inhibited the proliferation and colony-forming capacity of KKU100, KKU213A, and KKU213B cells. PA-CM also promoted the apoptosis of these CCA cells by causing the loss of mitochondrial membrane potential. Western Blotting confirmed that PA-CM induced CCA cell apoptosis by increasing the levels of the Bax/Bcl-2 ratio, cleaved caspase 3, and cleaved PARP, possibly by inhibiting the IL-6/JAK2/STAT3 signaling pathway. Moreover, our in vivo study also confirmed the suppressive effect of hPAMSCs on CCA cells by showing that PA-CM reduced tumor volume in nude mice transplanted with human CCA cells. Taken together, our results demonstrate that PA-CM has potent tumor-suppressive effects on human CCA cells and could potentially be used in combination with chemotherapy to develop a more effective treatment for CCA patients.

An injectable hydrogel based on MOF-derived hollow nanocomposites for starvation and gas therapy cooperated phototherapy

Here, glucose oxidase was loaded into Cu/Zn-MOF and then modified with polydopamine to obtain final nanoparticles (MGPs) with a size of approximately 150 nm. Under 808 nm laser irradiation, MGPs (0.125 mg mL−1) could rapidly increase 35.7 °C within 10 min, while generating large amounts of 1O2, and inducing starvation in the tumor microenvironment. L-arginine, as a precursor of NO, was incorporated into chitosan hydrogel together with MGP to allow for controlled release of NO under laser irradiation. Both in vitro and in vivo experiments demonstrated that an injectable hydrogel combining starvation and gas therapy with phototherapy effectively inhibited solid tumors growth.

Abstract C027: RSO-021, a first-in-class covalent inhibitor of mitochondrial PRX3: From bench to bedside

Abstract RSO-021 is a first-in-class mitochondrial peroxiredoxin 3 (PRX3) covalent small molecule inhibitor. The safety and tolerability of RSO-021 are currently being tested in the phase 1-2 MITOPE clinical trial in patients with malignant pleural effusion (MPE) arising from malignant mesothelioma or metastatic cancer to the lungs (NCT05278975). Altered redox status, resulting in elevated cellular hydrogen peroxide (H2O2) levels, drives the growth and proliferation of mesothelioma and other solid tumors. PRX3, the primary mitochondrial H2O2 scavenging enzyme, supports tumor cell escape from oxidative stress and aggressive tumor growth. RSO-021 attacks active site cysteine residues in PRX3 covalently to inactivate the enzyme, resulting in elevated H2O2 levels in the mitochondria incompatible with tumor cell survival. RSO-021 showed potent PRX3 inactivation and anti-tumor activity in pre-clinical cell and mouse models of mesothelioma with good tolerability supporting its clinical development. The MITOPE trial (NCT05278975) is an open-label, non-randomized, multicenter, translational phase 1-2 dose-escalation and expansion study designed to determine the safety, tolerability, dose-limiting toxicity (DLT), pharmacokinetics (PK), pharmacodynamics (PD), and preliminary anti-tumor activity of RSO-021 after intrapleural administration after complete drainage in patients with MPE due to mesothelioma or other metastatic disease. RSO-021 is administered locally as a pleural infusion through an indwelling catheter as a single dose on Day 1, 8 and 15 of each 3-week cycle. Safety is assessed at the end of the 21-day DLT period and efficacy every 6 weeks. PK and PD are assessed on pleural fluid, biopsies and with systemic sampling. RSO-021 is administered until disease progression, unacceptable toxicity, withdrawal of consent or study termination. Currently, MITOPE is actively recruiting patients in 4 open UK sites, with additional 9 sites on-boarding shortly. The dose escalation phase of the trial has 13 patients dosed at either 90, 120 or 180 mg flat dose levels and is anticipated to complete recruitment by the end of 2023. The trial will then continue to recruit patients as part of the dose expansion phase of the trial, which is exploring specific indications, including newly-diagnosed and previously-treated MM with MPE, as well as combination with systemic therapy in patients with refractory solid tumors and MPE. Citation Format: Dean A. Fennell, Sean Dulloo, Peter Szlosarek, Fiona Thistlethwaite, Simon Lord, Najib M Rahman, Brian Cunniff, Joanna Dzialo, Charlotte Poile, Rakesh Panchal, Jarrett Duncan, Pip Graham, Alice Bexon, George N. Naumov, James Spicer. RSO-021, a first-in-class covalent inhibitor of mitochondrial PRX3: From bench to bedside [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 C027.

Safety and Preliminary Efficacy of Sabestomig (AZD7789), an Anti-PD-1 and Anti-TIM-3 Bispecific Antibody, in Patients with Relapsed or Refractory Classical Hodgkin Lymphoma Previously Treated with Anti-PD-(L)1 Therapy

Background Immunotherapies targeting the PD-1/PD-L1 pathway have been shown to induce high response rates in patients with relapsed/refractory (r/r) classical Hodgkin lymphoma (cHL). However, most responses are partial and not durable. T cell immunoglobulin and mucin domain-containing protein-3 (TIM-3) is implicated as a resistance mechanism following anti-PD-1 therapy, and increased TIM-3 expression has been observed on T cells of patients with cHL post anti-PD-1 therapy. Therefore, we hypothesized that dual targeting of PD-1 and TIM-3 may reinvigorate immune responses and lead to more durable antitumor activity. Sabestomig is a monovalent, bispecific, humanized IgG1 monoclonal antibody that binds PD-1 and a unique epitope in the TIM-3 IgV domain without blocking phosphatidylserine binding, eliciting differential functionality. In preclinical mouse models, sabestomig was more effective at inhibiting growth of solid tumors than treatment targeting only PD-1, and sequential sabestomig after anti-PD-1 therapy increased antitumor responses. Here we report preliminary data from the ongoing dose escalation portion of a Phase I/II, open-label, multicenter study to assess the safety and preliminary efficacy of sabestomig in patients with r/r cHL (NCT05216835). Methods Eligible patients are aged ≥16 years with r/r cHL, an Eastern Cooperative Oncology Group performance status 0-1, at least one positron emission tomography-avid measurable lesion according to modified Lugano Criteria, and exposure to at least 2 prior lines of systemic therapy including a minimum of 3 cycles of an anti-PD-(L)1-based therapy. Sabestomig is intravenously infused over 1 hour following pretreatment with diphenhydramine and acetaminophen every 3 weeks, with planned doses ranging from 2 to 2000 mg across 8 cohorts (A1-A8); cohorts A1 to A4 (2, 7, 22.5 or 75 mg) follow an accelerated titration design with a single patient treated at each dose level, while cohorts A5 to A8 (225-2000 mg) follow a modified toxicity probability interval-2 algorithm. The primary endpoint of the dose escalation part of the study is safety, including dose-limiting toxicities (DLTs). Secondary endpoints include efficacy, pharmacokinetics, and immunogenicity. Data cutoffs were May 17, 2023 for safety and June 20, 2023 for efficacy. Results As of May 17, 2023, 14 patients were treated across the first 6 cohorts (A1-A6; 2-750 mg). They were predominantly male (64.3%) with Stage IV disease (85.7%) and a median age of 41.5 years (range, 25-77). Patients received a median of 6 prior lines of therapy (range, 3-13). Baseline disease characteristics are summarized in Table 1. Four patients received sabestomig 2-75 mg in cohorts A1-A4 (n=1 each), 5 patients received 225 mg in cohort A5, and 5 patients received 750 mg in cohort A6. Median duration of exposure to sabestomig was 8.9 weeks (range, 3.0-34.3) with a median of 3 cycles received (range 1-12). Sabestomig was well tolerated with no Grade ≥3 treatment-related adverse events (AEs) (Table 2). One Grade ≥3 AE occurred, which was fatal (sepsis secondary to gastric ulcer rupture, not related to sabestomig but deemed a DLT by the safety review committee). No immune-mediated AEs were reported, and no AEs led to treatment discontinuation. At the safety data cutoff, treatment was ongoing for 6 patients (42.9%). As of June 20, 2023, 11 patients had their first disease assessment and 4/11 had an objective response based on modified Lugano Criteria: 1 patient in cohort A3 (22.5 mg, a partial response [PR]) and all 3 patients who had their first disease assessment in cohort A6 (750 mg, 2 CRs and 1 PR). Two of these 4 responders (both in cohort A6) had experienced a best overall response of progressive disease during prior anti-PD-1 therapy; the other two initially had a response (1 CR [cohort A6] and 1 PR [cohort A3]) but subsequently progressed on anti-PD-1 therapy. Sustained PD-1 receptor occupancy (≥90%) was observed in peripheral blood at doses ≥225 mg. Conclusions Sabestomig was well tolerated with a manageable safety profile. Early efficacy data at 750 mg is encouraging with objective responses in 3/3 patients who had their first disease assessment, including those who were anti-PD-1 refractory or who relapsed on treatment. Updated data will be presented from ongoing and subsequent patients treated during dose escalation with doses of up to 2000 mg.

Development of a Novel Solid Tumor-Bearing Humanized Animal Model for Simultaneously Evaluating the Toxicity and Efficacy of Bispecific T Cell Engager

The development of novel therapeutics for cancer immunotherapy is tightly restricted by using human samples to estimate the drug toxicity before moving to clinical trials. Currently numerous tumor-bearing animal models have been utilized to test the efficacy of anti-tumor drugs. However, the success rate remains low in the development of clinical treatment. One prominent reason is that the utilized animal models could not accurately predict the toxic response of human immune system to the drugs. Thus, establishing a predictive nonclinical animal platform that can detect potential unwanted toxicity to human patient will be critical for improving the safety of immune drug development. Previously, we have demonstrated a disseminated tumor-bearing humanized mouse model that can evaluate the toxicity and efficacy of bispecific T cell engager (BiTE). Here we aim to develop a novel tumor-bearing mouse model that can simultaneously evaluate the long-term efficacy against solid tumor in addition to cytokine level assessment. In this study, we co-engrafted disseminated and solid tumor cells to the same mice and compared with mice engrafted with disseminated tumor or solid tumor. Three donors were tested. Method: Human triple negative breast cancer MDA-MB-231/Luciferase-2A-GFP (MDA) cell line is epidermal growth factor receptor (EGFR) positive and is labeled with luciferin. The cells were implanted into PBMC humanized NSG-MHC class I/II double knock out (DKO) mice via orthotopic injection into mammary fat pad (MFP, 5x10 6 cells/mouse) and/or via intravenous (IV, 1x10 6/mouse) injection. Disseminated MDA cells normally colonize lung of mice after IV injection. Following EGFRxCD3 BiTE dosing initiation, tumor burden in MFP and lung were evaluated by a in vivo Xenogen imaging system, and solid tumor volume in MFP was measured 2 to 3 times a week by a digital caliper. To evaluate the toxicity levels in response to EGFRxCD3 BiTE treatment, the level of inflammatory cytokine levels in serum of mice were analyzed by BD cytometric bead array. Clinical observations, body weight change, and human immune cell activation were also monitored for drug toxicity assessment. Results: EGFRxCD3 BiTE treatment induced elevated human inflammatory cytokines in serum from humanized mice implanted with combined MFP and IV tumor. By comparison, the serum cytokine levels were low in mice implanted with MDA cells via MFP alone, suggesting that IV injection of tumor cells is required for cytokine production induced by EGFRxCD3 BiTE. MFP solid tumor continued to grow in the PBS control group and EGFRxCD3 BiTE treatments significantly inhibited solid tumor growth regardless the presence of IV implanted with MDA cells. Conclusion: Without disseminated tumor, MFP solid tumor-bearing mice served as a model for efficacy but did not produce inflammatory cytokines after treatment with EGFRxCD3 BiTE. Combination of solid and disseminated tumor implantation provides a new tumor inoculation approach that can be used for simultaneously evaluating the toxic cytokine release and long-term efficacy of immunotherapy drugs. This is a promising preclinical platform to define optimal approaches for clinical cancer treatment.

Polysaccharide of Asparagus cochinchinensis (Lour.) Merr: Preliminary characterization and antitumor activity in vivo

A novel high molecular weight polysaccharide (ACP) extracted and purified from Asparagus cochinchinensis (Lour.) Merr was preliminarily characterized by HPGPC, Ion chromatography, FT-IR, NMR, and SEM, and its in vivo antitumor activity was investigated using an H22 tumor-bearing mice model. ACP was identified to be a homogeneous component with a purity of 95.75 ± 1.58%, a molecular weight of 74.7 kDa, and an acidic polysaccharide composed of Rha, Ara, Gal, Glc, Xyl, Man, GalA, and GlcA at a molar ratio of 0.34:1.86:8.72:15.99:1:0.52:2.25:0.11 with α- and β-anomeric units. In vivo antitumor activity evaluation showed that ACP could effectively inhibit the growth of solid tumors in H22 hepatocellular carcinoma-transplanted mice at inhibition rates of 26.5% and 45.3%, respectively, with the two possible apoptosis mechanisms involving arresting the cell cycle in G0/G1 phase and reducing the mitochondrial membrane potential in H22 cells. These results indicate the potential of ACP as a therapeutic agent of liver cancer.

Targeting isoforms of RON kinase (MST1R) drives antitumor efficacy.

Recepteur d'origine nantais (RON, MST1R) is a single-span transmembrane receptor tyrosine kinase (RTK) aberrantly expressed in numerous cancers, including various solid tumors. How naturally occurring splicing isoforms of RON, especially those which are constitutively activated, affect tumorigenesis and therapeutic response, is largely unknown. Here, we identified that presence of activated RON could be a possible factor for the development of resistance against anti-EGFR (cetuximab) therapy in colorectal cancer patient tissues. Also, we elucidated the roles of three splicing variants of RON, RON Δ155, Δ160, and Δ165 as tumor drivers in cancer cell lines. Subsequently, we designed an inhibitor of RON, WM-S1-030, to suppress phosphorylation thereby inhibiting the activation of the three RON variants as well as the wild type. Specifically, WM-S1-030 treatment led to potent regression of tumor growth in solid tumors expressing the RON variants Δ155, Δ160, and Δ165. Two mechanisms for the RON oncogenic activity depending on KRAS genotype was evaluated in our study which include activation of EGFR and Src, in a trimeric complex, and stabilization of the beta-catenin. In terms of the immunotherapy, WM-S1-030 elicited notable antitumor immunity in anti-PD-1 resistant cell derived mouse model, likely via repression of M1/M2 polarization of macrophages. These findings suggest that WM-S1-030 could be developed as a new treatment option for cancer patients expressing these three RON variants.

Bisabosqual A: A novel asparagine synthetase inhibitor suppressing the proliferation and migration of human non-small cell lung cancer A549 cells

Asparagine synthetase (ASNS) is a crucial enzyme for the de novo biosynthesis of endogenous asparagine (Asn), and ASNS shows the positive relationship with the growth of several solid tumors. Most of ASNS inhibitors are analogs of transition-state in ASNS reaction, but their low cell permeability hinders their anticancer activity. Therefore, novel ASNS inhibitors with a new pharmacophore urgently need to be developed. In this study, we established and applied a system for in vitro screening of ASNS inhibitors, and found a promising unique bisabolane-type meroterpenoid molecule, bisabosqual A (Bis A), able to covalently modify K556 site of ASNS protein. Bis A targeted ASNS to suppress cell proliferation of human non-small cell lung cancer A549 cells and exhibited a synergistic effect with L-asparaginase (L-ASNase). Mechanistically, Bis A promoted oxidative stress and apoptosis, while inhibiting autophagy, cell migration and epithelial-mesenchymal transition (EMT), impeding cancer cell development. Moreover, Bis A induced negative feedback pathways containing the GCN2-eIF2α-ATF4, PI3K-AKT-mTORC1 and RAF-MEK-ERK axes, but combination treatment of Bis A and rapamycin/torin-1 overcame the potential drug resistance triggered by mTOR pathways. Our study demonstrates that ASNS inhibition is promising for cancer chemotherapy, and Bis A is a potential lead ASNS inhibitor for anticancer development.