bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heterofunctional polymeric degrading chimeras (HYDRAC) for targeted protein degradation.
Authors: Wang Max Mu , Truica Mihai Ioan , Gattis Brayley , Abdulkadir Sarki , Gianneschi Nathan 
Published: June 2023

e15106 Background: Here we utilize a novel proteomimetic polymer platform technology which packages peptide side chains as globular structures with benificial emergent properties for the development of compounds capable of targeted protein degradation. We refer to this class of compounds as Heterofunctional polYmeric DegRAding Chimeras (HYDRACs) and demonstrate the selective degradation of the oncoprotein Myc as a proof-of-concept. Methods: HYDRACs were generated using ring-opening metathesis polymerization (ROMP), resulting in structures highly resistant to proteolysis. Heterofunctional constructs were designed incorporating secondary sequences in addition to Myc-targeting H1. The degron sequence (RRRG) was incorporated to engage endogenous cellular machinery for targeted protein degradation. Immunofluorescence/confocal microscopy were used to assess cellular penetration; cellular viability assays coupled with use of both cellular (isogenic pairs, Myc-dependent and independent lines) and biochemical (scrambled H1 sequence, homopolymers, monomeric peptides) controls were used to assess tumor antiproliferative effects and Myc-specific responses. Myc target engagement was assessed via immunoprecipitation assays, NMR, circular dichroism, and fluorescence polarization with protein degradation monitored by Western blot. A CRISPR knockdown library screen of all known E3 ligases and mass spec was used to identify pathways responsible for HYDRAC-induced protein degradation. Results: A library of HYDRACs with narrow polydispersity and predetermined degrees of polymerization were generated. These compounds are cell-penetrating with antiproliferative effects at micromolar IC50s. HYDRAC treatment resulted in significant decreases in Myc protein levels, which was rescued by inhibition with MG132 and MLN4924. RNAseq showed selective overexpression of Myc pathway genes. Biophysical analysis showed strong HYDRAC-Myc interactions, supported by the presence of Myc protein following pull-down of biotin-terminated HYDRACs. Identification of E3 ligases implicated in the degradation pathway using a combination of a CRISPR knock down library and mass spectrometry are ongoing. Mice bearing Myc-CaP tumors showed delayed tumor growth following treatment with Myc-degrading HYDRACs. Studies on biodistribution and pharmacokinetic profile of the compound, as well as the ability to degrade other proteins of interest using combinations of degron sequences are ongoing. Conclusions: The presented work demonstrates the feasibility of HYDRACs for targeted protein degradation of Myc, providing rationale for further studies in Myc-driven tumor models. This successful demonstration of this first-in-class compound also opens the door for the development of HYDRACs targeting other proteins of interest and incorporation of different E3 ligase recruiters for tissue or disease-specific activity.
Droplet digital PCR as an alternative method for detecting MET amplification in patients with non-small cell lung cancer.
Authors: Su Jun-wei , Weng Cheng-Di , Yang Jin-Ji 
Published: June 2023

e15107 Background: Mesenchymal-epithelial transition (MET) amplification is associated with resistance to epidermal growth factor receptor tyrosine kinase inhibitors in lung cancer patients and poor prognosis. Fluorescence in situ hybridization (FISH) is the most commonly used method for detecting MET amplification but is not possible in cases where tissue samples are unavailable. Here, we evaluated the utility of droplet digital PCR (ddPCR) in detecting MET amplification in non-small cell lung cancer (NSCLC) patients. Methods: We evaluated a c-Met: EFTUD ddPCR assay of the c-Met gene and using EFTUD as the reference gene to determine c-Met amplification in plasma cfDNA samples from 80 advanced NSCLC patients. A total of 84 plasma samples were collected for MET amplification detection using ddPCR. Paired tissue samples using FISH were to evaluate the correlation between the MET-ddPCR ratio and MET FISH. Results were correlated with clinical findings. Results: We found that ddPCR had an 88.1% concordance rate with FISH in detecting MET CNG, with an area under the curve (AUC) of 0.84. Eleven patients were positive for MET amplification by FISH prior to MET inhibitor treatment, and these patients had a partial response rate (PR) of 45.5% and a median progression-free survival (PFS) of 12.1 months. However, MET amplification identified by ddPCR failed to distinguish significant clinical outcomes. The PR rate and median PFS for ddPCR-positive vs. negative patients were 57.1% vs. 50.0% and 12.1 months vs. 2.1 months (P = 0.276). Conclusions: Our results suggest that ddPCR is a reliable alternative to detect MET amplification in plasma cfDNA samples. Met status determined in cfDNA may have potential as a predictive factor for PFS after c-Met inhibitor therapy.
Cardiovascular adverse events associated with second generation Bruton tyrosine kinase inhibitor therapy: A systematic review and meta-analysis.
Authors: Proskuriakova Ekaterina , Jasaraj Ranjit , Gaire Suman , Raut Anuradha , Reddy Vijay Ketan , Shrestha Dhan , Kovalenko Iuliia , Khosla Paramjeet Grewal 
Published: June 2023

e15109 Background: Cardiovascular adverse events (CVAE) is a frequent adverse effect of first-generation bruton tyrosine kinase inhibitor (BTKi) that significantly reduce the usage of these medications. This led to the development of second-generation BTKi (acalbrutinib and zanubrutinib) that are more selective and potent with presumably better safety profile than first-generation. CVAE associated with second-generation BTKis in the clinical practice has been sporadically reported. Hence, we attempted to pool the reported CVAE in this meta-analysis. Methods: Systematic searches of PubMed, PubMed Central, Cochrane library for clinical trials, Scopus, and Embase were conducted from inception until January 14, 2023. This meta-analysis performed per PRISMA protocol and registered in PROSPERO database (CRD42023392406). All randomized clinical trials including acalabrutinib or zanabrutinib in treatment arm with standard of care as control arms were included. The literature found from search engines were imported in the covidence platform and screened independently by 2 reviewers in title/abstract and full text phase. Conflicts were resolved by a third reviewer. Included studies were thoroughly reviewed by all reviewers and data from them extracted in a Microsoft Excel. Analysis performed using RevMan 5.4 meta-analysis software. Odds ratio was used as an effect measure with 95% confidence intervals (CI) using random effect model. Results: A total of ten trials (with 3048 patients, 1710 in treatment arm) were included in this meta-analysis. Cardiovascular events leading to death were reported in 4 studies. In the second generation BTKis group cardiovascular death was 0.94% (7/744), while in the control group it was 1.7% (12/705). Using the random effect model, a pooled analysis on death due to cardiovascular events were statistically insignificant across two groups (OR 0.46, 95% CI 0.19 to 1.11; n = 2039; I2 = 57%). Similarly, pooled analysis for any bleeding/hemorrhage from eight studies (OR 2.12, 95% CI 0.91 to 4.97; n = 2611; I2 = 93%), major bleeding from seven studies (OR 1.12; 95% CI 0.66 to 1.89, n = 2658, I2 = 28%), acute coronary syndrome from four studies (OR 3.13; 95% CI 0.65 to 15.13, n = 1585, I2 = 0%) were also statistically not significant. Conclusions: Treatment with acalabrutinib and zanabrutinib did not show any significant differences in cardiovascular events between the group when compared with standard treatment as a control group, meta-analyzing the data from published trials. As there is no evidence of worse cardiovascular outcomes or superiority of the second-generation BTKis compared to standard treatments in terms of safety based on available evidence, further large-scale controlled trials are required to support that new-generation BTKis are superior in safety and better tolerated.
Examination of in vitro and in vivo antitumor activity of Menin-MLL inhibitor, D0060-319.
Authors: Xu Huan , Dai Yazhao , Li Li , Zhu Hui 
Published: June 2023

e15119 Background: Rearrangements of the mixed-lineage leukemia (MLL) gene result in the production of MLL fusion proteins, which occurs in 5-10% of acute leukemia in adults and > 70% of infant leukemia, respectively. Menin protein, encoded by the MEN1 gene, interacts with MLL fusion proteins to induce leukemia. D0060-319, a Menin-MLL inhibitor, shows a significant antitumor effect in vivo and in vitro. Methods: The fluorescence polarization (FP) binding assay to assess inhibition of D0060-319 to Menin-MLL interaction. The cross reactivity of 44 molecular targets was tested to assess their off-target binding potential. The anti-proliferative ability of D0060-319 was evaluated by CCK-8 in different cell lines. The in vivo anti-tumor effect of D0060-319 was evaluated in a human MV4-11 and MOLM-13 cell mouse xenograft tumor model. Briefly, five million MV4-11 cells or 500 thousand MOLM-13 cells were subcutaneously implanted the right flank of CB-17 SCID mice to establish cell mouse xenograft tumor model. When tumor volume reached 100-120mm3, the animals were grouped and administrated with D0060-319 at the dose of 25, 50, 75, 100 mg/kg or vehicle at twice daily for 21days. Tumor measurements and body weights were performed twice weekly to assess the antitumor efficacy and tolerability of D0060-319. Results: D0060-319 competitively binds Menin to inhibit the interaction with MLL fusion protein with IC50 of 7.46nM, has high antiproliferative activity in MV4-11 and MOLM-13 cell lines containing MLL fusion proteins with IC50 values of 4.0nM and 1.7nM, respectively. However, in Kasumi-1, K562, HL-60 and KG-1 cell lines lacking MLL rearrangement, D0060-319 antiproliferative activity with IC50 > 10 μM, indicating selectivity for leukemia cell lines containing MLL fusion proteins active. In the MV4-11 cell mouse xenograft tumor model, D0060-319 significantly inhibited tumor growth at the dose of 25mg/kg, with a tumor growth inhibition (TGI) rate of 82.97%, and other doses (50, 75 and 100mg/kg) tumor completely regressed. In the MOLM-13 cell mouse xenograft tumor model, a dose-dependent anti-tumor effect was observed after 14 days of administration, with TGIs of 81.08%, 88.47% and 90.26% at the dose of 50, 75 or 100 mg/kg, respectively. In both CDX models, no bodyweight loss was observed, suggesting that D0060-319 was well tolerated. Furthermore, off-target screening assays showed that D0060-319 did not cross-react with any of the 44 molecular targets at 10 μM, indicating that D0060-319 is highly selective, high-affinity for the MLL-binding pocket on Menin. Conclusions: In MLL-rearranged leukemia, D0060-319 showed high antitumor activity and good tolerability both in vitro and in vivo.
Phase 1, randomized, open-label, single-dose, crossover study to evaluate the bioequivalence of four formulations of oral rivoceranib tablets in healthy subjects.
Authors: Weyer Janelle , Meng Xianzhang , Finis Lynda , Strickland Bill , Park CheolHee , Jang Seong 
Published: June 2023

e15122 Background: Rivoceranib is a novel oral tyrosine kinase inhibitor that potently and selectively inhibits VEGFR2. Rivoceranib is being investigated for indications targeted towards solid malignancies as either monotherapy or in combination with other anticancer therapies. Herein, we assessed the bioequivalence of a single dose of rivoceranib administered as 4 different formulations in healthy subjects. Methods: This single-center, open-label, randomized, single-dose, 4-way crossover study evaluated the bioequivalence of 4 different formulations of rivoceranib oral tablets in healthy adults. Each subject participated in 4 treatment periods, where they were randomized to 1 of 4 sequences: ABCD, BDAC, CADB, and DBAC (Formulation A = rivoceranib 250 mg tablet/clinical formulation used in the pivotal phase 3 study, Formulation B = rivoceranib 200 mg tablet/clinical formulation used in early clinical studies, Formulation C = rivoceranib 250 mg tablet/formulation to be developed for future use, Formulation D = rivoceranib 250 mg tablet/to-be-marketed formulation). Results: Of the 60 subjects enrolled, 66.7% were male, 88.3% were white, and median age was 43 years. The median plasma rivoceranib Tmax was similar following all treatments (2 hours post-dose). The 90% CIs around the geometric mean ratios (GMRs) of plasma rivoceranib AUC0-t, AUC0-inf, and Cmax for Formulation B vs. Formulation A and Formulation D vs. Formulation A were within the 80-125% reference interval, demonstrating bioequivalence between Formulation B and Formulation A as well as Formulation D and Formulation A. The 90% CIs around the GMRs of plasma rivoceranib AUC0-t, AUC0-inf, and Cmax for Formulation C vs. Formulation A were slightly outside of the 80%-125% reference interval. Conclusions: Formulations B (clinical formulation used in early clinical studies) and D (to-be-marketed formulation) were bioequivalent to Formulation A (clinical formulation used in the pivotal phase 3 study). Formulation C (formulation to be developed for future use) and Formulation A were similar, but the difference was slightly outside of the bioequivalence criteria. It remains to be evaluated whether the difference in bioavailability between Formulation C and Formulation A is clinically meaningful. Clinical trial information: NCT05287360 . [Table: see text]
Preclinical efficacy of NXP900, a YES1/SRC kinase inhibitor in esophageal squamous cancer models.
Authors: Dash Sweta , Nyswaner Katherine , Harvey Mungo J.B. , King Ben , Woods Allison , Carragher Neil O. , Unciti Asier , Poradosu Enrique , Brognard John 
Published: June 2023

e15133 Background: NXP900 (eCF506) is a novel potent and selective SRC family kinase (SFK) inhibitor, (IC50 of 0.47 nM against YES1). NXP900 locks its target into its native “closed” conformation (type 1.5 inhibitor), thereby inhibiting both kinase activity and complex formation with protein partners (Temps et al. Cancer Res. 2021, 81, 5438-5450). In contrast, multi-kinase inhibitors, including dasatinib and bosutinib, block SRC in the active “open” conformation (type 1 inhibitors) promoting the association of SFK and signaling partners via allosteric facilitation of conformational protein activation (Higuchi et al. Cell Rep 2021, 34, 108876). YES1 gene amplification is prevalent in several squamous cancers including esophageal squamous cell carcinoma (ESCC), lung SCC, head and neck SCC, and bladder cancer, indicating that YES1 would be an attractive target in these cancer types. Moreover, crosstalk between YES1 and the Hippo pathway suggests that NXP900 may have a therapeutic potential in squamous cancers with Hippo pathway alterations. Methods: Animal strain: CD1 Nude mice (Charles River). Human ESCC cancer cell lines: KYSE70, OE21, KYSE410, KYSE30 and OE19 cells (Sigma), TE5, and TE14 (Riken). Xenograft tumors were generated by subcutaneous implantation on the right lower flank of the thigh at a cell density of 2x106 to 1x107 cells/mouse at 0.1 mL Matrigel/cell dilution volume per injection. Experiment groups: Vehicle, NXP900 (40 mg/kg); treatment: QD for 28 days, oral gavage. Colony formation assay: 500 cells of 4 cell lines (KYSE70, OE21, KYSE410 and KYSE30) and 1500 cells of 3 cell lines (OE19, TE5 and TE14) were seeded in 24 well plates in triplicates; and treated with NXP900 for 14 days followed by staining with crystal violet. Results: NXP900 strongly inhibited cell proliferation of KYSE70, KYSE410, KYSE30, OE21, OE19, TE5 and TE14 in a colony formation assay. NXP900 significantly inhibited the activating phosphorylation of SRC in KYSE70 cells at 10 nM, while no effect was observed on the inhibitory tyrosine residues on YES1/FYN at 250 nM. Oral administration of NXP900 in a KYSE70 xenograft mouse model with YES1 gene amplification, resulted in an average decrease of tumor volume of 71% while in the vehicle control group tumor volume increased by 472% (p ≤ 0.001), demonstrating significant tumor regression. On Day 27, a mean increase from baseline in body weight of 2.0% and 3.5% was seen in the NXP900 and control groups respectively, indicating good tolerability. Conclusions: NXP900 is a novel potent and selective YES1/SRC kinase inhibitor with subnanomolar IC50 against YES1. NXP900 has marked antitumor efficacy in esophageal carcinoma models in vitro and in vivo providing substantial proof of concept for targeting solid tumors with YES1 and Hippo pathway alterations. IND enabling studies for NXP900 have been completed.
Effects of namodenoson on pancreatic carcinoma: Preclinical evidence.
Authors: Fishman Pnina , Bareket-Samish Avital , Itzhak Inbal 
Published: June 2023

e15134 Background: Namodenoson, an A3 adenosine receptor (A3AR) agonist, is currently in a phase 3 trial for the treatment of advanced liver cancer. The mechanism of action of namodenoson involves de-regulation of the Wnt and NF-κB signaling pathways, followed by an increase in pro-apoptotic proteins and Fas-ligand, resulting in tumor growth inhibition. As the Wnt signaling pathway is also highly implicated in pancreatic carcinogenesis, we examined the anti-growth effect of namodenoson on pancreatic carcinoma cell lines and investigated the molecular mechanism involved. Methods: BxPC-3 human pancreatic carcinoma cells were cultured in the presence and absence of 0.01, 0.1, and 1 nM namodenoson. A combined treatment of namodenoson (0.1 nM) and gemcitabine (0.2 µM) was also studied. 3[H]-thymidine proliferation and MTT assays were used to monitor cell growth, and Western blot analyses were performed to identify the involved regulatory cell growth proteins. Results: Results of the 3[H]-thymidine proliferation assays demonstrated significant dose-dependent inhibition of the growth of BxPC-3 cells with namodenoson (1 nM: 67.4% ± 1.7%, p < 0.001; 0.1 nM: 53.7% ± 6.3%, p < 0.05; 0.01 nM: 27.9 % ± 2.3%, p < 0.005). The MTT results revealed that a combined treatment with namodenoson plus gemcitabine had an additive inhibitory effect (namodenoson: 48.6% ± 1.4%; gemcitabine: 44.4% ± 0.7%; namodenoson plus gemcitabine: 65.4% ± 1.4%; p < 0.001 for all). Western blot analyses showed that namodenoson treatment was associated with downregulation of the Wnt pathway regulatory proteins including p-Akt, NF- κB, GSK-3β, and β-catenin. Conclusions: Our findings showed that nanomolar concentrations of namodenoson inhibit the growth of pancreatic carcinoma via A3AR activation and de-regulation of the Wnt/β-catenin pathway, both as a monotherapy and in combination with gemcitabine. Thus, these results support a potential role for namodenoson in treating pancreatic cancer, thereby opening a novel therapeutic opportunity for this disease.
Programmed cell death ligand 1 (PD-L1) expression landscape and its relationship with tumor mutational burden (TMB) in a community-based cancer center patient population in rural central Nebraska.
Authors: Copur Mehmet Sitki , Peterson Tonya , Doornbos Kathryn , Tun Soe Min , Springer Carlene R. , Robbins Leslie , Arbogast Jessica , Muske Chandra , Buescher Lisa , Sukup Jordan , Buescher Lisa , Horn Adam , Wedel Whitney , Lintel Nicholas , Brunt Kathryn , Duckert Randall , Lavudi Swathi 
Published: June 2023

e15141 Background: PD-L1 and TMB are the most widely used immunotherapy biomarkers to identify populations who would benefit from immunotherapy, with the higher values predicting better therapeutic efficacy. We sought to explore the occurrence and correlation of these two biomarkers in a community based cancer center patient population in rural central Nebraska. Methods: One hundred sixty consecutive cancer patients(pts) diagnosed and treated between 2018-2022 at Morrison Cancer Center were evaluated. Malignant tumor tissue specimens were analyzed for PD-L1expression and TMB (Foundation test). PD-L1 expression was classified as low ( < 1%), intermediate (1% ≥ PD-L1 < 50%), high (≥ 50%), and TMB classified as low [1–5 mutations/Megabase (muts/Mb)], intermediate (6–19 muts/Mb), high (≥ 20 muts/Mb)]. PD-L1 and TMB values were treated as continuous variables to calculate a Pearson correlation coefficient and p value. Results: Mean age at diagnosis was 64 years (range 33-89). Malignant histology diagnoses included 55 pts with lung, 18 colorectal, 12 breast, 15 unknown primary, 15 pancreas, 10 renal, 3 melanoma, 1 thyroid, 6 ovary, 5 uterus, 5 head and neck, 5 prostate, 5 biliary, and 1 vulvar. Fifty four percent (86/160) of pts had low, 33% (53/160) had intermediate, and 13% (21/160) had high PD-L1 expression score with a mean PD-L1 value of 13.8% (range 0-100). Sixty six percent (106/160) of pts had low, 30% (48/160) had intermediate, and 4% had high TMB with a mean TMB value of 5.6 muts/Mb (range 0-66). A marginal linear correlation between PD-L1 expression and TMB was observed (Pearson Correlation Coefficient = 0.172, P = 0.0270). Conclusions: PD-L1 and TMB are both important biomarkers for immunotherapy. Increasing availability and use of both PD-L1 and TMB testing in the community oncology practice beseeches the question of frequency of these two biomarkers in various cancers in the community oncology patient population as well as any association between them. The results of our study gives a glimpse of the distribution landscape of these two biomarkers for community oncology patients and suggest that PD-L1 and TMB may be independent markers with a weak association. PD-L1 and TMB may each inform use of immunotherapy by different mechanisms. [Table: see text]