Therapeutic Modalities Research Articles

Targeting the receptor tyrosine kinase MerTK shows therapeutic value in gastric adenocarcinoma.

Despite multiple therapeutic modalities, the overall survival of patients with gastric adenocarcinoma remains poor, especially for advanced tumor stages. Although the tyrosine kinase MerTK has shown therapeutic relevance in several tumor entities, its potential effects in gastric adenocarcinoma have not yet been sufficiently characterized. MerTK expression and its influence on patient survival were evaluated by immunohistochemistry in a cohort of 140 patients with gastric adenocarcinoma. CRISPR/Cas9 knockout and siRNA knockdown of MerTK in the gastric cancer cell lines SNU1, SNU5, and MKN45 was used to analyze protein expression, growth, migration, and invasion properties invitro and in a murine xenograft model. MerTK was pharmacologically targeted with the small molecule inhibitor UNC2025 invitro and invivo. In patients, high MerTK expression was associated with decreased overall survival (OS) and lymph node metastasis especially in patients without neoadjuvant therapy (p < 0.05). Knockout and knockdown of MerTK reduced cell proliferation and migration both invitro and invivo. UNC2025, a small-molecule inhibitor of MerTK, exhibited a significant therapeutic response invitro and invivo. Additionally, MerTK expression attenuated the response to neoadjuvant treatment, and its inhibition sensitized tumor cells to 5-Fluorouracil (5-FU)-based chemotherapy invitro. Our findings demonstrate the potential value of MerTK as a prognostic biomarker for gastric adenocarcinoma. Targeting MerTK may become a new treatment option, especially for patients with advanced tumors, and may overcome resistance to established chemotherapies.

Role of biologics in severe food allergy.

This review examine the dynamic landscape of food allergy treatment within the context of emerging biologics. Our purpose is to comprehensively evaluate the potential benefits, challenges, and transformative impact associated with the utilization of biologics in comparison to conventional therapeutic modalities. This document synthesizes recent scientific investigations to various biologics, such as omalizumab, ligelizumab, dupilumab, and tezepelumab, providing a nuanced understanding of their roles in oral immunotherapy, rapid desensitization, and overall food allergy management. Recent studies and clinical trials highlight the impact of anti-IgE treatment on food allergies, revealing critical findings such as dose-related efficacy, facilitation of rapid desensitization in peanut allergies, and the sustained positive outcomes observed in individuals with multifood allergies. The use of biologics presents a groundbreaking approach in the treatment of food allergies. The multifaceted action of these agents, along with their potential to overcome the challenges associated with traditional therapies, marks a significant advancement. Despite the persisting challenges of economic constraints and the need for further safety studies, biologics offer a promising avenue for improving the quality of life for individuals with food allergies. Ongoing research and collaborative efforts are imperative to fully realize the transformative potential inherent in these emerging therapeutic frontiers.

Activity of Matrix Metalloproteinase-7 (MMP-7) Related to Heparin-Binding Epidermal Growth Factor (HB-EGF) Activation in Squamous Cell as Diagnostic Biomarker for Early Detection And Treatment of Maxilarian Cancer: Review Article

Background: Squamous cell carcinoma (SCC) has a higher chance of occurance and progression in maxillary sinus cancer than other paranasal sinus cancers (80%). Matrix metalloproteinase-7 (MMP-7) is a protease specified in collagen type IV degradation and Heparin Binding Epidermal Growth Factor activation, which causes an increase in squamous cell carcinoma proliferation and metastasis Methods: This review aims to identify the potential role of MMP-7 as a target for prevention and therapeutic modality to manage SCC sinus maxillary cancer. We did literature searching in several databases to elucidate the importance of the MMP-7 pathway in SCC sinus maxillary cancer progression. Results: : Increasing expression and activity of MMP-7 are associated with maxillary sinus cancer progression. Squamous cells, the most impacted in maxillary sinus cancer, sustain dysplasia due to heparin-binding epidermal growth factor (HB-EGF) activation into EGF by MMP-7 and Cluster of Differentiation 44 (CD44) binding complexes that trigger high proliferation and mitogenic activity. The therapeutic function of MMP-7 occurs by affecting the protein 53 (p53) in cancer cell apoptosis initiation by specific interaction with CD44. Conclusion: MMP-7 could be an alternative therapeutic target and potential treatment option for maxillary sinus cancer. Furthermore, more trials should be done to test the relevancy of MMP-7 uses as an SCC sinus maxillary cancer modality

HighFold: accurately predicting structures of cyclic peptides and complexes with head-to-tail and disulfide bridge constraints.

In recent years, cyclic peptides have emerged as a promising therapeutic modality due to their diverse biological activities. Understanding the structures of these cyclic peptides and their complexes is crucial for unlocking invaluable insights about protein target-cyclic peptide interaction, which can facilitate the development of novel-related drugs. However, conducting experimental observations is time-consuming and expensive. Computer-aided drug design methods are not practical enough in real-world applications. To tackles this challenge, we introduce HighFold, an AlphaFold-derived model in this study. By integrating specific details about the head-to-tail circle and disulfide bridge structures, the HighFold model can accurately predict the structures of cyclic peptides and their complexes. Our model demonstrates superior predictive performance compared to other existing approaches, representing a significant advancement in structure-activity research. The HighFold model is openly accessible at https://github.com/hongliangduan/HighFold.

Management of pancreatic and duodenal trauma in childhood: a university hospital experience over a 10-year period.

Duodenal/pancreatic injuries occur in less than 10% of intra-abdominal injuries in pediatric blunt trauma. Isolated duodenal/pancreatic injuries occur in two-thirds of cases, while combined injuries occur in the remaining. This study aimed to investigate pediatric patients with pancreatic and duodenal trauma. Data from 31 patients admitted to Atatürk University, Medical Faculty, Department of Pediatric Surgery for pancreatic/duodenal trauma between 2010 and 2019 were retrospectively analyzed. Age/gender, province of origin, duration before hospital admission, trauma type, injured organs, injury severity, diagnostic and therapeutic modalities, complications, hospitalization duration, blood transfusion requirement, and mortality rate were recorded. Twenty-four patients were male, and 7 were female. The mean age was 9years. The leading cause was bicycle accidents, with 12 cases, followed by traffic accidents/bumps, with 7 cases each. Comorbid organ injuries accompanied 18 cases. Duodenal trauma was most commonly accompanied by liver injuries (4/8), whereas pancreatic injury by pulmonary injuries (7/23). Serum amylase at initial hospital presentation was elevated in 83.9% of the patients. Thirty patients underwent abdominal CT, and FAST was performed in 20. While 54.8% of the patients were conservatively managed, 45.2% underwent surgery. Because of the anatomical proximity of the pancreas and the duodenum, both organs should be considered being co-affected by a localized trauma. Radiologic confirmation of perforation in duodenal trauma and an intra-abdominal pancreatic pseudocyst in pancreatic trauma are the most critical surgical indications of pancreaticoduodenal trauma. Conservative management's success is increased in the absence of duodenal perforation and cases of non-symptomatic pancreatic pseudocyst.

Tailoring Treatment in Cardiovascular Diseases: The Role of Targeted Therapies.

Cardiovascular diseases (CVDs) remain the leading cause of morbidity and mortality around the globe. To address this public health burden, innovative therapeutic agents are being developed to specifically target molecular and genetic markers. Various therapeutic modalities have been implemented, including vaccines, monoclonal or bispecific antibodies, and gene-based therapies. Such drugs precisely target the underlying disease pathophysiology, aiming at notable molecules such as lipid metabolism regulators, proinflammatory cytokines, and growth factors. This review focuses on the latest advancements in different targeted therapies. It provides an insightful overview of the current landscape of targeted cardiovascular therapies, highlighting promising strategies with potential to transform the treatment of CVDs into an era of precision medicine.

Chemical Peels for Melasma: A Systematic Review.

Melasma is a common chronic, relapsing pigmentary disorder that causes psychological impact. Chemical peels are a well-known therapeutic modality used for accelerating the treatment of melasma. To review the published evidence on the efficacy and safety of chemical peels in the treatment of melasma. A systematic review was done. A meta-analysis could not be done due to the heterogeneity of data. The authors conducted a PubMed search and included prospective case series of more than 10 cases and randomized controlled trials (RCTs) that have studied the safety and/or efficacy of chemical peel in melasma. Out of 24 studies, 9 were clinical/comparative trials and 15 were RCTs. The total sample size was 1,075. The duration of the study varied from 8 to 36 weeks. Only 8 studies were split face. All studies used self-assessment, physician global assessment, and Melasma Area and Severity Index (MASI) for quantifying the results. Glycolic acid was found to be the most safe and effective in melasma. Chemical peels were found to be safe and effective in the management of melasma.

Experimental and Computational Analysis of High-Intensity Focused Ultrasound Thermal Ablation in Breast Cancer Cells: Monolayers vs. Spheroids.

High-intensity focused ultrasound (HIFU) is a non-invasive therapeutic modality that uses precise acoustic energy to ablate cancerous tissues through coagulative necrosis. In this context, we investigate the efficacy of HIFU ablation in two distinct cellular configurations, namely 2D monolayers and 3D spheroids of epithelial breast cancer cell lines (MDA-MB 231 and MCF7). The primary objective is to compare the response of these two in vitro models to HIFU while measuring their ablation percentages and temperature elevation levels. HIFU was systematically applied to the cell cultures, varying ultrasound intensity and duty cycle during different sonication sessions. The results indicate that the degree of ablation is highly influenced by the duty cycle, with higher duty cycles resulting in greater ablation percentages, while sonication duration has a minimal impact. Numerical simulations validate experimental observations, highlighting a significant disparity in the response of 2D monolayers and 3D spheroids to HIFU treatment. Specifically, tumor spheroids require lower temperature elevations for effective ablation, and their ablation percentage significantly increases with elevated duty cycles. This study contributes to a comprehensive understanding of acoustic energy conversion within the biological system during HIFU treatment for 2D versus 3D ablation targets, holding potential implications for refining and personalizing breast cancer therapeutic strategies.

Single-Center Experience of Pediatric Cystic Kidney Disease and Literature Review.

Pediatric cystic kidney disease (CyKD) includes conditions characterized by renal cysts. Despite extensive research in this field, there are no reliable genetics or other biomarkers to estimate the phenotypic consequences. Therefore, CyKD in children heavily relies on clinical and diagnostic testing to predict the long-term outcomes. A retrospective study aimed to provide a concise overview of this condition and analyze real-life data from a single-center pediatric CyKD cohort followed during a 12-year period. Medical records were reviewed for extensive clinical, laboratory, and radiological data, treatment approaches, and long-term outcomes. During the study period, 112 patients received a diagnosis of pediatric CyKD. Male patients were more involved than female (1:0.93). Fifty-six patients had a multicystic dysplastic kidney; twenty-one of them had an autosomal dominant disorder; fifteen had an isolated renal cyst; ten had been diagnosed with autosomal recessive polycystic kidney disease; three had the tuberous sclerosis complex; two patients each had Bardet-Biedl, Joubert syndrome, and nephronophthisis; and one had been diagnosed with the trisomy 13 condition. Genetic testing was performed in 17.9% of the patients, revealing disease-causing mutations in three-quarters (75.0%) of the tested patients. The most commonly presenting symptoms were abdominal distension (21.4%), abdominal pain (15.2%), and oligohydramnios (12.5%). Recurrent urinary tract infections (UTI) were documented in one-quarter of the patients, while 20.5% of them developed hypertension during the long-term follow-up. Antibiotic prophylaxis and antihypertensive treatment were the most employed therapeutic modalities. Seventeen patients progressed to chronic kidney disease (CKD), with thirteen of them eventually reaching end-stage renal disease (ESRD). The time from the initial detection of cysts on an ultrasound (US) to the onset of CKD across the entire cohort was 59.0 (7.0-31124.0) months, whereas the duration from the detection of cysts on an US to the onset of ESRD across the whole cohort was 127.0 (33.0-141.0) months. The median follow-up duration in the cohort was 3.0 (1.0-7.0) years. The patients who progressed to ESRD had clinical symptoms at the time of initial clinical presentation. This study is the first large cohort of patients reported from Croatia. The most common CyKD was the multicystic dysplastic kidney disease. The most common clinical presentation was abdominal distention, abdominal pain, and oliguria. The most common long-term complications were recurrent UTIs, hypertension, CKD, and ESRD.

Efficacy of Theta Burst Stimulation in Bipolar Depression: A Systematic Review and Meta-analysis

Purpose: Repetitive transcranial magnetic stimulation (rTMS) has emerged as a promising alternative therapeutic modality for bipolar depression. Theta Burst Stimulation (TBS) a novel variant of rTMS is currently evolving as an effective and safe therapeutic tool in the management of bipolar depression, although it is still in the initial phase. In the present systematic review and meta-analysis, data from various studies investigating the effectiveness and tolerability of TBS in individuals with bipolar depression were synthesized. Data collection and analyses: Eight studies assessing the efficacy of TBS as a treatment for bipolar depression were included after a systematic search. The quality of the included studies was assessed using standard methods. Synthesized data included three uncontrolled clinical trials (non-randomized controlled trials [RCTs]; n = 48) and five RCTs ( n = 201). While four studies used intermittent TBS, two studies used continuous TBS and bilateral TBS, each. Data from the five RCTs meta-analyzed using the random effects model showed the pooled odds ratio and effect size for response in the active group compared to the sham group were 1.01 (95%CI: 0.56, 1.83) and 1.64 (95%CI: –0.46, 3.74), respectively. Although both models showed no/minimal heterogeneity or publication bias, they were not statistically significant. A supplementary trial sequential analysis confirmed the need for more studies and suggested that the total sample required for drawing meaningful inferences was 918 samples. Conclusion: The available evidence suggests that more studies are required to draw conclusive inferences for the efficacy of TBS in the therapeutic management of bipolar depression.

Human Herpes Virus-6 (HHV-6) Reactivation after Hematopoietic Cell Transplant and Chimeric Antigen Receptor (CAR)- T Cell Therapy: A Shifting Landscape.

HHV-6B reactivation affects approximately half of all allogeneic hematopoietic cell transplant (HCT) recipients. HHV-6B is the most frequent infectious cause of encephalitis following HCT and is associated with pleiotropic manifestations in this setting, including graft-versus-host disease, myelosuppression, pneumonitis, and CMV reactivation, although the causal link is not always clear. When the virus inserts its genome in chromosomes of germ cells, the chromosomally integrated form (ciHHV6) is inherited by offspring. The condition of ciHHV6 is characterized by the persistent detection of HHV-6 DNA, often confounding diagnosis of reactivation and disease-this has also been associated with adverse outcomes. Recent changes in clinical practice in the field of cellular therapies, including a wider use of post-HCT cyclophosphamide, the advent of letermovir for CMV prophylaxis, and the rapid expansion of novel cellular therapies require contemporary epidemiological studies to determine the pathogenic role and spectrum of disease of HHV-6B in the current era. Research into the epidemiology and clinical significance of HHV-6B in chimeric antigen receptor T cell (CAR-T cell) therapy recipients is in its infancy. No controlled trials have determined the optimal treatment for HHV-6B. Treatment is reserved for end-organ disease, and the choice of antiviral agent is influenced by expected toxicities. Virus-specific T cells may provide a novel, less toxic therapeutic modality but is more logistically challenging. Preventive strategies are hindered by the high toxicity of current antivirals. Ongoing study is needed to keep up with the evolving epidemiology and impact of HHV-6 in diverse and expanding immunocompromised patient populations.

Abstract 3545: Atlas of therapeutic targets: Evolution of druggable proteome

Abstract Selection of therapeutic targets and modalities is of fundamental importance in drug discovery, as it can have a direct impact on the economics of drug discovery as well as its success rate. With the advancement of chem-bio and proteomics methods together with advent of artificial intelligence, the scope of the druggable proteome is expanding. The choice of drug targets gains further importance as we link it to potential clinical efficacy and safety, genetic variation and patient populations. Drug targets are often selected based on inadequate understanding of target biology, tractability by existing toolbox of therapeutic modalities and insufficient insight into possible clinical outcome. We have developed an “Atlas of Therapeutic Targets”, an interactive resource that provides classification of drug target families with linked information on clinical and investigational drugs. The atlas also provides key insights and score for tractability of targets with various therapeutic modalities. Particularly, our scoring system grades the targets on dimensions such as ligandability, involvement in protein-protein complexes, structural disorder, hotspot analysis and cross-referencing with known targets, thus yielding a total tractability score of 1-10. Dissection of targets based on such dimensions, enables critical evaluation therapeutic modalities as well tools and methodologies needed for their conversion into successful drug discovery programs. We curate 900+ human drug targets, analysis of which indicates the continued dominance of privileged target families across disease areas, but also the growth of novel first-in-class mechanisms. This atlas will be available as a resource for the wide drug discovery community. Citation Format: Ivan Babic, Elmar Nurmemmedov. Atlas of therapeutic targets: Evolution of druggable proteome [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 3545.

Abstract 5684: The role of the lncRNA Slamon and Slc25a13 neighbor gene in melanoma aggressiveness

Abstract Melanomas are responsible for about 80% of all deaths related to skin cancers. This is due to its high potential of metastasizing and developing resistance to treatments. Novel therapeutic modalities and the identification of biomarkers able to predict the prognosis are necessary. Studies have revealed the central role of long non-coding RNAs (lncRNAs) in regulatory networks controlling cell behavior. The disruption of these regulatory networks as a consequence of altered expression of lncRNAs can contribute to cancer development and progression. Our laboratory has developed a linear cellular model of melanoma progression consisting of distinct cell lines: melan-a (melanocytes), 4C (pre-malignant melanocytes), 4C11- (undifferentiated, slow-growing and non-metastatic melanoma cells) and 4C11+ (differentiated, highly proliferative and metastatic melanoma cells), which were analyzed for their lncRNA expression profile. Differentially expressed lncRNAs identified by RNAseq were selected by in silico analyses based on their proximity to differentially expressed coding-genes in the same melanoma model. Correlation analyses were performed between the expression of differentially expressed lncRNA neighboring genes and their melanoma patient survival, using two independent melanoma cohorts (TCGA and Leeds). Among those lncRNA located near genes presenting prognostic value is the lncRNA Gm20619, here named Slamon, neighbor to Slc25a13 gene (mitochondrial aspartate/glutamate carrier gene), which high expression correlates with poor prognosis. Both Slamon lncRNA and Slc25a13 are highly expressed in the metastatic melanoma cells 4C11+ compared to melan-a, 4C and 4C11- cell lines. The knocking down of Slamon or Slc25a13 were performed in 4C11+ melanoma cells, followed by analyses of collective migration, clonogenicity, proliferation, anoikis resistance, and dacarbazine and MEK inhibitor treatment. By knocking down Slamon lead 4C11+ cells significantly less migratory and less resistant to dacarbazine, and resulted in increased expression of Slc25a13 gene. 4C11+ melanoma cells knocked down for Slc25a13 also presented reduced migratory, clonogenicity, proliferation and anoikis resistance capability, diminished sensitivity to dacarbazine and MEK inhibitor treatment, and increased expression of Slamon lncRNA. The analysis of genes coexpressed with Slc25a13 revealed genes related to tumor progression, as Cdk15, Lsp1 and Pdgfrb. Our results suggest a crucial role of the lncRNA Slamon and Slc25a13 coding gene in melanoma progression which has never been described before. Slamon regulates tumor aggressiveness associated with Slc25a13, forming a regulatory network. These findings reveal the importance of lncRNAs in melanoma biology, potentially indicating transcripts that can serve as prognostic biomarkers and therapeutic targets in melanoma. Citation Format: Beatriz C. Tonin, Ana L. Ayub, André H. Lengert, Sang W. Han, Eduardo M. Reis, Frank J. Slack, Miriam G. Jasiulionis. The role of the lncRNA Slamon and Slc25a13 neighbor gene in melanoma aggressiveness [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 5684.

Abstract 5814: EPriL macrocycles as a platform for the rapid generation of effective kinase degrader antibody conjugates (DACs)

Abstract The therapeutic success of antibody drug conjugates (ADCs) drives a continuous search for novel payloads that can increase therapeutic window and thereby widen the applications for ADCs. Recently, heterobifunctional degraders have gained great interest as payloads, and degrader antibody conjugates (DACs) are seen as a novel therapeutic modality. Heterobifunctional degraders consist of a small molecule ligand that binds a target (protein of interest or POI), a spacer and an E3 ligase ligand, which can catalyze target degradation. Owing to their catalytic activity, degraders can have better potency than the equivalent inhibitors, making them suitable as ADC payload [1]. Degrader payloads can target a wider variety of mechanisms than classic payloads, which use the same antitumor strategies as chemotherapy, such as tubulin binding or topoisomerase inhibition. Since many FDA-approved targeted therapies are based on the inhibition of protein kinases, we investigated degraders of these kinases as DAC payloads. Kinase DACs could bring enhanced targeting and therefore better therapeutic window to a field where classic ADC payloads have often shown substantial toxicities. As many heterobifunctional degraders show poor cell membrane penetration, their inhibitory potential could be increased as part of a DAC, where endocytic uptake is followed by intracellular release of the degrader payload. To identify kinase degrader payloads, we present a workflow based on a platform called Energetically Privileged Ligands (EPriLs). EPriLs are macrocycle scaffolds that bind non-covalently in the kinase ATP pocket. Their unique binding mode avoids contacts with amino acid positions where resistance to kinase inhibitors frequently occurs. EPriL macrocycles can be decorated appropriately to rationally design specific inhibitors for many therapeutically relevant kinases, and provide synthetic handles to couple them to VHL or CRBN ligands to generate effective kinase degraders. Here we describe how EPriL kinase degraders can be developed into effective DACs, using consecutive libraries of EPriL ligands, spacers, E3 ligase ligands and linkers. First, suitable degraders are identified, based on rapid and deep target degradation and potent antiproliferative activity on target cell lines. Degraders are then transformed into maleimide-linked degraders using convenient attachment of enzymatically cleavable linkers. In a medium throughput fashion, these maleimides are coupled to antibodies to generate DACs, which are tested for stability and biological potency. Applying this workflow to various well-validated kinase targets in oncology resulted in a promising kinase targeting DAC with favorable ADME properties, clear potentiation compared to the parent degrader, and increased selectivity for tumor cell lines. 1] Dragovich et al., Chem. Soc. Rev. (2022) 51, 3886-3897. Citation Format: Joost C. Uitdehaag, Jos e Man, Michelle Muller, Freek an Cauter, Sander an Gemert, Milan Hoffmann, Yvonne G. an Mil, Winfried R. Mulder, Martine B. Prinsen, Jan Gerard Sterrenburg, Diep Vu, Joeri e Wit, Erik Ensing, Rogier C. Buijsman. EPriL macrocycles as a platform for the rapid generation of effective kinase degrader antibody conjugates (DACs) [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 5814.

Abstract 4484: Discovery of a potential first-in-class Za targeted small molecule inhibitor of ADAR1 p150 which demonstrates strong anti-tumor efficacy

Abstract Background: Adenosine deaminase, RNA specific (ADAR1), catalyzes the hydrolytic deamination of adenosine (A) to inosine (I) in double-stranded (ds) RNAs. ADAR1 has two isoforms: p110 in the nucleus and p150 in the cytoplasm. Both isoforms alter the coding and non-coding sections of self-dsRNA. The ADAR1 p150 isoform is expressed from an interferon (IFN)-response promoter and has a Z-DNA/Z-RNA binding domain at the N-terminus. ADAR1 has been implicated in promoting cell survival and resistance to immune response in several cancer types like multiple myeloma, breast, lung, liver, and skin cancer. Inhibition of ADAR1 p150 has the potential to enhance anti-tumor efficacy as monotherapy and in combination with other therapeutic modalities. The Zα domain of ADAR1 p150 provides exclusivity to p150 compared to the other isoforms. Herein, we outline the discovery of a potential first-in-class Zα targeted ADAR1 p150 inhibitor and degrader for cancer immunotherapy. Methods: AVA-ADR-001 was identified through a high throughput p110 knockout cell-based assay. The ability of AVA-ADR-001 to induce interferons was confirmed in various cell lines. The anti-tumor efficacy of AVA-ADR-001 was evaluated in B16F10 syngeneic melanoma mice model as monotherapy and in combination with anti-PD-1. PROTAC AVA-ADR-703 was used to demonstrate selective degradation of the ADAR1 p150 protein. Results: We have discovered AVA-ADR-001, a novel small molecule inhibitor of ADAR1 which significantly increases the IFN response in vitro in an MDA5-dependent manner. In vitro binding studies confirm that AVA-ADR-001 binds to the Zα domain of ADAR1 with low nM potency. AVA-ADR-001 demonstrates anti-tumor efficacy 1.5X superior to anti-PD1 ICB non-responsive tumor models. In addition, the AVA-ADR-001 monotherapy and combination groups showed a significant upregulation of numerous interferon-stimulated genes, including IFIH1, IFN-β, and CXCL-10. Finally, targeted protein degradation of ADAR1 p150 while sparing ADAR1 p110 was confirmed when cells were treated with AVA-ADR-703 with nM DC50, the PROTAC variant of AVA-ADR-001. Conclusions: To our knowledge, AVA-ADR-001 and AVA-ADR-703 are the first selective Zα-targeted small molecule inhibitor and degrader of ADAR1 p150 respectively to be disclosed. As a powerful and targeted first-in-class ADAR1 inhibitor, AVA-ADR-001 has demonstrated a strong induction of interferon (IFN) in a number of cancer cell lines and in vivo. AVA-ADR-001 has also demonstrated strong in vivo anti-tumor efficacy as monotherapy and in combination with anti-PD1. Furthermore, AVA-ADR-703 demonstrates selective and highly efficient degradation of ADAR1 p150 while sparing ADAR1 p110. Given that ADAR1 has a pro-metastatic and immune-suppressive role, AVA-ADR-001 and AVA-ADR-703 have significant potential to emerge as strong cancer immunotherapy candidates. Citation Format: Avijit Goswami, Sandeep Goyal, Kawaljit Singh, Princy Khurana, Barnali Deb, Aditya Kulkarni. Discovery of a potential first-in-class Za targeted small molecule inhibitor of ADAR1 p150 which demonstrates strong anti-tumor efficacy [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 4484.

Abstract 6220: TRIP13 augments pancreatic cancer progression- An integrated systems biology study

Abstract Background- Pancreatic cancer (PanCa) is one of the most aggressive forms of cancer and its incidence rate is continuously increasing every year. It is expected that by 2030, PanCa will become the 2nd leading cause of cancer related deaths in the United States due to the lack of understanding of the complexity of disease, early diagnostic methods, and extremely poor survival. Despite great advancements in biomedical research, there are very limited modalities available for the early detection of PanCa and its treatment. Thus, understanding of disease biology and identification of newer diagnostic and therapeutic modalities are high priority research in the field. Our group has identified Thyroid Receptor Interacting Protein -13 (TRIP13) as an oncogenic protein which contributes to early events of PanCa. Methodology- High dimensional omics and structural elucidation of TRIP13 was conducted using publicly available databases like Consurf, Phosphosite, GTEx and GEPIA2. Molecular biology techniques were applied to cross validate all our bioinformatics data. qPCR, immunoblotting and IHC analysis was performed in progressive PDAC cell lines and human tissues to decipher the expression level of TRIP13 in various pathological staging including functional enrichment analysis using Linkdomics. Results- The structural elucidation analysis revealed that most of the functionally exposed residues, phosphorylation and ubiquitylation sites situated at AAA domain of TRIP13. As compared to normal, TRIP13 is significantly over expressed in pancreatic tumor samples. Pathological staging 2 and 1 (early stages) showed relatively higher expression of TRIP13 as compared advanced stage. Of the total 7 isoforms of TRIP13, only one isoform (TRIP13-001) has shown the over expression in pancreatic tumor samples and shown association with poor survival. In correlation studies, TRIP13 has shown positive association with other pancreatic cancer biomarkers (CEACAM5, S100A4, MUC1, MSLN, CA125). The functional enrichment analyses suggest that TRIP13 is involved in important patho-physiological pathways like DNA repair, viral carcinogenesis, cellular senescence, and cell cycle. Our wet lab experiments also support our computational biology observation. Conclusion- This integrated computational biology study suggests a potential role of TRIP13 in pancreatic cancer progression via modulation of important oncological pathways. Wet lab experimentations also validated the involvement of TRIP13 in pancreatic cancer progression. TRIP13 expression may be associated with patient prognosis. Thus, its inhibitory small molecules can be useful for boosting pancreatic cancer therapies in clinical settings. Key words- TRIP13, Pancreatic cancer, Early events of cancer, Integrative Biology, Transcriptomics, Computational biology Citation Format: Swati Dhasmana, Anupam Dhasmana, Stella Rios, Iris A. Perez, Sheema Khan, Farrukh Afaq, Upender Manne, Murali M. Yallapu, Subhash C. Chauhan. TRIP13 augments pancreatic cancer progression- An integrated systems biology study [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 6220.

NIR‐II Excitable Semiconducting Polymers with AIE Characteristics for Fluorescence‐Photoacoustic Imaging‐Guided Synergistic Phototherapy

AbstractThe development of semiconducting polymers (SPs) with simultaneous second near‐infrared (NIR‐II) absorption and multimodal phototheranostic functions is highly desired in the field of oncotherapy. Aggregation‐induced emission luminogens (AIEgens) have been acknowledged to possess unique potential in integrating multiple diagnostic and therapeutic modalities into one organic molecule. Nevertheless, SPs are hard to achieve AIE activity due to their extended π‐conjugated skeletons as well as the large and planar configuration of commonly‐used electrophilic acceptor moieties. Herein, an ingenious acceptor dimerization‐based acceptor distortion strategy is proposed in current work by taking advantage of the sterically encumbered acceptors with large and planar structure for the successful construction of AIE‐active SPs. Through further finely adjusting the molecular donor–acceptor interaction strength, one of the obtained AIE‐active SPs named SP3 bearing the highest intramolecular charge transfer effect presented desired NIR‐II absorption and aggregation‐induced NIR‐II fluorescence emission, good reactive oxygen species production ability, as well as superior photothermal conversion performance. Accordingly, SP3 is selected to fabricate into nanoparticles and successfully applied in the NIR‐II laser‐triggered fluorescence‐photoacoustic imaging‐guided photodynamic‐photothermal therapy of tumors. This study represents the first NIR‐II excitable AIE‐active SP, and offers a new perspective on designing advanced multimodal phototheranostic polymers for efficient treatment of malignant tumor.

Abstract 2110: Characterization of the correlation between BTK degradation and tumor growth inhibition of the BTK target protein degraders using PK/PD modeling

Abstract Introduction: Chimeric degradation activation compound (CDACs) are heterobifunctional molecules causing target protein degradation by simultaneously binding to the target protein as well as an E3-ubiquitin ligase. As an emerging new therapeutic modality, it is critical to understand the contribution of compound-specific parameters (e.g. drug exposure and binding potency to the target protein) and the system-specific parameters (e.g. target protein turnover) of CDACs to its in vivo effect. Method: To identify the key parameters for guiding compound optimization and to find the correlation between BTK degradation and the extent of tumor growth inhibition, a mechanistic PK/PD model was built (Phoenix WinNonlin 8.1) on a dataset generated from a BTK CDAC (Compound A). The PK/PD model was simplified mathematically compared to the full mechanistic models published previously1, since the binding affinity of Compound A to BTK was much higher than the binding affinity of CDAC to E3 ligase. This model can be linked to a tumor growth inhibition (TGI) mathematical model to find the correlation between target degradation and efficacy.Result: In mouse Rec-1 xenograft model, the BTK degradation and TGI was studied. The mice were dosed with Compound A orally at 0.3, 1, or 3 mg/kg, dose-dependent PK and BTK degradation in tumor was observed. After a single dose administration of Compound A, ~50% BTK degradation was observed in tumor at 24 h post dosing and the tumor BTK rebound to baseline at 72 hours post dosing at 3 mg/kg. The tumor BTK degradation was deeper after repeated dosing. A PK/PD model was built based on the data in this PK/PD study. The calculated BTK turnover half-life in Rec-1 model is about 16 h. In the efficacy study, Compound A was dosed orally once a day to the mice at 0.3, 1, or 3 mg/kg. The 3 mg/kg group achieved ~90% tumor growth inhibition in Rec-1 xenograft model. The PK/PD model was then linked with a mathematical model describing TGI2. A threshold level of BTK degradation in tumor to achieve tumor stasis was identified to be 96% for Rec-1 tumor model. At 90% tumor growth inhibition, average of 70% BTK was predicted to be degraded in tumor at steady state. The model was subsequently used to characterize the PK/PD relationship of BGB-16673. The efficacy of BGB-16673 was predicted using the model and it agreed with the observed value.Conclusions: The current model deepened our understanding of the PK/PD relationship of CDACs. The approach can be used to simulate different scenarios, e.g. different BTK turnover, potency or PK exposure. It can also be used in compound selection and optimization to predict repeated dose PD and efficacy from a single dose PK/PD data.1. J Pharmacokinet Pharmacodyn. 2021 Feb;48(1):149-163.2. Cancer Res. 2004 Feb 1;64(3):1094-101. Citation Format: Yue Wu, Shuran Li, Fan Wang, Nan Hu, Longbo Yin, Xiaomin Song, Jiye Zhang, Aiying Xu, Shasha Yang. Characterization of the correlation between BTK degradation and tumor growth inhibition of the BTK target protein degraders using PK/PD modeling [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 2110.

Abstract 2106: Creating a living public oncology drug resource using PRISM cell line viability screening

Abstract Cancer treatment options are rapidly expanding, with many new targeted agents and therapeutic modalities under development. However, systematic characterization of the cellular effects of emerging drug candidates is limited. Screening such agents against the approximately 900 diverse cancer cell lines in the PRISM assay can provide insights into drug specificity and cancer subtype selectivity and, leveraging the rich genomic and functional characterization of PRISM cell lines, support for annotated mechanisms of action. Our goal was to generate a large, high-quality, oncology-focused dataset that can benefit the cancer research community by characterizing new test agents of high clinical relevance. In the past year, this effort has been utilized to screen a total of 180 test agents with plans to extend testing to an additional 140 over the next year. The collection includes biologics (e.g., cytokines, antibodies, antibody-drug conjugates), targeted protein degraders, and small molecules. Supporting the quality of the screening collection and the robustness of the PRISM assay, the majority of inhibitors (including mutant-selective KRAS inhibitors, and inhibitors targeting the WRN helicase, a preferential vulnerability in cancers with microsatellite instability) tested to date closely mimic the effects of genetic knockout and/or knockdown of their annotated target, as measured by CRISPR and shRNA data available through the Dependency Map Portal (depmap.org). Further, while we observe strong modality-independent co-clustering of test agents annotated to act on the same target(s) (e.g., tucatinib with the antibody-drug conjugates T-DM1 and T-DXd for HER2), intriguing differences in efficacy and selectivity emerge in individual cell line models. All PRISM reference screening data and analyses will be made publicly available through depmap.org to serve as a benchmark for new drug development candidates and to further the efforts of researchers worldwide to match emerging cancer therapies to the patients most likely to benefit. Citation Format: Matthew G. Rees, Mustafa Kocak, Claudine Mapa, Dennie Frederick, Ellen Nguyen, Alvin Kalathungal, Anthony Fazio, John F. Davis, Emily Reeves, Ariah Tucci, Rachael Barry, Colleen Harrington, Jillian Eskra, Melissa M. Ronan, William R. Sellers, Jennifer A. Roth. Creating a living public oncology drug resource using PRISM cell line viability screening [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 2106.

Abstract 5315: Bispecific antibody discovery platform for IND enabling studies: Case study of HER-2/NKG2D bispecific antibody

Abstract There has been a surge in the number of therapeutic antibody modalities primarily attributable to the increasing array of antibody engineering platforms. Among these, the emergent class of bispecific antibodies has exhibited remarkable potential compared to conventional monoclonal antibodies, marked by their targeting precision, expanded therapeutic scope, and personalized medicine prospects. Presently, eight bispecific antibodies are approved, with over 100 in varying clinical trial phases. Using a bispecific targeting HER-2 and NKG2D, we outline a comprehensive end-to-end platform for bispecific antibody discovery and characterization, designed to facilitate the progression of novel bispecific antibodies through the antibody discovery pipeline. To secure the most optimal binders tailored to specific targets, our methodology capitalizes on an extensive repertoire of phage display libraries. This strategy is instrumental in the identification of antibody candidates with exceptional binding affinity, broad cross-species compatibility, and enhanced drug-like properties. From the anti-HER2 chimeric antigen receptor ML39 and anti-NKG2D Fab KYK2.0, we synthesized four distinct bispecific formats by fusing single-chain variable fragments (scFv) to immunoglobulin G (IgG) molecules. These formats varied in terms of valency and scFv positioning, allowing us to assess their impact on antibody expression, binding kinetics, and functional efficacy. Subsequently, the bispecific antibodies underwent comprehensive in vitro efficacy and safety characterization, employing multiple innovative technologies. Utilizing a distinctive label-free approach with the xCELLigence system, we elucidated the kinetics of immune cell-mediated target cell killing upon antigen-antibody complex recognition in specific cancer cells. Safety assessments, with a focus on on-target/off-tumor effects, were conducted using the Retrogenix platform. Furthermore, cytokine profiling was utilized to evaluate potential risks associated with enhanced cytokine levels, reinforcing the specificity and safety profile of the bispecific antibody. Finally, pertinent in vivo models, including peripheral blood mononuclear cells (PBMC) and immune cell transfer models, facilitated the translation of preclinical bispecific antibody evaluations into clinically relevant contexts. This comprehensive platform for bispecific antibody discovery and functional characterization serves as a robust foundation for supporting Investigational New Drug (IND) applications, thus facilitating the transition of novel bispecific antibodies from the pre-clinical to the clinical phase of the drug discovery process. Citation Format: Laura Ratcliffe, Rachel Pooley, Katherine Carr, Hamed Janbazacyabar, David Cobeta Lopez, Deborah Bruce, Sarah L. Martin, Matthew Benson, Jonathan White, Benita Quist, Sophie Vermond, Gemma Moiset, Sophie Hill, Renny Feldman, Paul Wan, Eva Oswald, Folkert Verkaar, Mark Aspinall-O'Dea, Julia Schueler, Namrata Jayanth. Bispecific antibody discovery platform for IND enabling studies: Case study of HER-2/NKG2D bispecific antibody [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 5315.