bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of cost of Oncotype DX versus CanAssist Breast, an Indian prognostic test, in the management of node-negative early breast cancer in a single centre in India.
Authors: Bapna Ajay , Samar Aseem Kumar , Gupta Tarachand , Agarwal Pawan
Published: August 2023

14

Background: It is well known that prognostication is important as not all women benefit equally from adjuvant chemotherapy, especially in HR+/HER2- early breast cancer (BC). Multiple publications have showcased the cost-effectiveness of Oncotype DX (ODX) in various countries. However, ODX remains out of reach for many patients in under-resourced geographies like Asia, Africa etc where healthcare insurance is not a norm. CanAssist Breast (CAB) is an affordable prognostic test developed on Indian patients. CAB couples the immunohistochemistry technique with an AI/ML-based prediction model to segregate patients as either low-risk or high-risk for distant breast cancer recurrence. CAB has been validated in the USA, Spain, Germany, Austria, Italy, and The Netherlands for predicting 5–10-year distant recurrences and has comparable low-risk accuracy with that of ODX/MammaPrint. In this abstract, we present real-world data about cost-analysis of the use of CanAssist Breast versus ODX in a single cancer care centre in Rajasthan, a state in Western India that has a per capita income of $ 1504 as against $ 68,400 in the USA. Methods: 124 patients with node-negative early BC were treated at a trust hospital in Jaipur, Rajasthan from 2020-2022. All the high-risk patients received chemotherapy at the treating clinician’s discretion. For the cost analysis model, the cost of chemotherapy drugs, hospitalization charges, and costs involved in managing short-term adverse events was compared for CAB and ODX. Treatment is provided at subsidized costs to the patients in this trust hospital. Results: The median age of this cohort was 55 years (35-78), with primarily T2 and G2 diseases. High-risk proportions were 27% and 17% (as per TAILORx) and by CAB and ODX respectively. The cost of ODX is 5.8 times higher than the cost of CAB. The cost savings per patient with CAB over ODX were US $ 3477 (Rs. 278161) in this trust hospital. Conclusions: This economic model shows that the use of ODX could impose an economic burden due to its cost. Use of CAB would lead to savings without comprising the patient’s health, as both tests have comparable performance.
Differential therapeutic benefit of platinum-doublet chemotherapies with ICIs based on smoking status plus PD-L1 expression in advanced NSCLC patients.
Authors: Fukuda Akito , Yoshida Tatsuya , Masuda Ken , Shinno Yuki , Okuma Yusuke , Goto Yasushi , Horinouchi Hidehito , Yamamoto Noboru , Ohe Yuichiro
Published: August 2023

148

Background: Chemotherapy combined with immune checkpoint inhibitors (Chemo-ICIs) has been the standard of care for advanced NSCLC patients. Smoking status is associated with PD-L1 expression and the TILs within tumor microenvironment, which leads to the efficacy of ICIs. It remains unclear how the combined with PD-L1 expression and smoking status affect the efficacy of Chemo-ICIs. Methods: We retrospectively reviewed advanced NSCLC patients without driver mutation (EGFR, ALK, ROS-1, BRAF) who received Chemo-ICIs from January 2019 to June 2021, and Chemo from January 2014 to December 2016 as a first-line therapy at the National Cancer Center Hospital, Tokyo, Japan. The additional benefit of ICIs to chemotherapy were evaluated by comparing the Chemo-ICIs group with Chemo group. Results: During study period, 225 patients received Chemo-ICIs and 133 received Chemo as a first line therapy. Overall response rate (ORR) and progression free survival (PFS) was significantly better in the Chemo-ICIs group than the Chemo group (ORR: 44.9% vs. 40.2%, p=0.01, and median PFS: 8.2 months vs. 6.2 months, P<0.001). The median overall survival was 27.9 months in Chemo-ICIs and 20.4 months in chemo (P=0.14). In the subgroup analysis of PFS, the benefit of Chemo-ICIs compared to Chemo was not observed in never-smoker patients (8.9months vs 8.8months; HR 0.72; 95% CI, 0.49 to 1.28; P=0.27). In contrast, among smoker patients, Chemo-ICIs showed significant longer PFS compared to Chemo (7.5 months vs 5.7 months, P<0.001). Next, patients received Chemo-ICIs were classified into six groups, based on the smoking status (never smoker or smoker) and PD-L1 expression (TPS: 0%, 1-49%, or 50%) (Table). Chemo-ICIs showed better PFS rate at 12 months in the smoker/ TPS ≥50% group (42.2% vs 25.0%, p=0.03), and the never smoker/TPS ≥50% group (75.0% vs 27.3%, p=0.05) compared with Chemo (Table). In patients with 1-49% TPS, compared to the Chemo group, significant PFS benefit of ICIs was observed only in smoker patients (42.2% vs 25.0%, p=0.003), not in never smoker patients (46.2% vs 27.3%, p=0.13). In patients with 0% TPS, although the additional benefit of ICIs to Chemo was not observed both in never smoker (13.3% vs 27.3%, p=0.47) and smoker patients (26.6% vs 25.0%, p=0.13), PFS rate at 12 months in never smoker (13.3%) was lowest among all six groups. Conclusions: Our study suggested that additional benefit of ICIs to chemotherapy could be predicted by combined with smoking status and PD-L1 expression. [Table: see text]
Retrospective observational study of outcomes in HER2-positive metastatic breast cancer patients treated with trastuzumab emtansine in single center.
Authors: Hsieh WanChen , Chang Chia lun
Published: August 2023

15

Background: Trastuzumab emtansine (T-DM1) was established as a standard second line treatment option for metastatic HER2 positive breast cancer (mBC). However, the EMILIA study did not include disease progression patients who received pertuzumab plus trastuzumab previously. After CLEOPATRA study, the dual blockade treatment was indicated for advanced breast cancer as first line treatment. In this study, we attempted to build the T-DM1 predictive model and to evaluate the association between dual blockade and T-DM1 for metastatic HER2-positive breast cancer. Methods: This retrospective cohort study was conducted in Taiwan. Consecutive patients with metastatic HER2-positive breast cancer who had received T-DM1 between January 2013 and December 2022 were enrolled. Those patients were divided to two groups based on strategy of trastuzumab only or dual-blockade as prior therapy. Demographic, clinical, and pre- and post-T-DM1 treatment characteristics were described. The swimmer plots were used to estimate time to treatment discontinuation (TTD) and overall survival (OS). Results: Of 32 patients treated with T-DM1, 12 (38%) had prior treatment with pertuzumab plus trastuzumab. The median age was 60 years. Most patients had visceral disease and 59% had two or more prior treatments for mBC before T-DM1 (range 1–8 lines). Most patients (63%) stopped T-DM1 due to disease progressed and followed by financial problem (22%). In dual-blockade group, median TTD with T-DM1 was 4.5 months; median OS from diagnosis day was 68.2 months (8 patients still alive in the end of study). In trastuzumab only group, median TTD with T-DM1 was 3.8 months; median OS from diagnosis day was 73.2 months (5 patients still alive in the end of study). In our swimmer plots we found a trend that the shorter interval between trastuzumab +/- pertuzumab and T-DM1 the longer TTD with T-DM1. Conclusions: Patients treated with T-DM1 in this study appeared to have more advanced disease than patients in clinical trials and were treated in later lines of therapy. We should prescribe T-DM1 once trastuzumab with or without pertuzumab failure for the longer TTD. The small patient number was a major limitation in our study. A multi-center analysis is needed and it will help us for treatment suggestion under the Taiwan’s National Health Insurance clinical practice.
Furmonertinib as adjuvant therapy in postoperative EGFR-mutated stage IA2-IIIA non-small cell lung cancer (NSCLC) with high-risk pathological factors.
Authors: Wu Ziheng , Zhang Qingyi , Ke Lei , He Tianyu , Lv Wang , Hu Jian
Published: August 2023

150

Background: NSCLC patients with high-risk pathological factors such as micropapillary (MP), solid (S), spread through air spaces (STAS), and visceral pleural invasion (VPI) have shown poor prognosis in previous studies. Furmonertinib (AST2818) is a novel, promising oral third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), which has demonstrated efficacy in tumours harbouring sensitive EGFR mutations and T790M resistance mutation. Here, we investigated the efficacy and safety of furmonertinib as adjuvant therapy in postoperative EGFR-mutated NSCLC patients with high-risk pathological factors. Methods: Patients who underwent radical lung cancer surgery with both EGFR mutations and MP/S/STAS/VPI were enrolled and received furmonertinib 80mg daily. The adjuvant therapy time (6-36months) depended on patients’ pathologic stage and physical conditions. The disease-free survival (DFS), safety and tolerability were evaluated. Results: This study retrospectively analyzed 69 patients who were pathologically confirmed adenocarcinoma, EGFR mutation positive (exon 19 deletion/L858R/exon 20ins), stage IA2-IIIA NSCLC. All of them had at least one high-risk pathological factor. There were 44 (63.8%) patients with MP, 14 (20.3%) with S, 10 (14.5%) with STAS and 33 (47.8%) with VPI. All patients were followed at least 13 months, and 24 (34.8%) of them have been followed up for over 21 months, median follow-up time was 18.5 months. By the cut-off date of March 1, 2023, all patients were alive and with no radiographic recurrence, including 3 (4.3%) with exon 20ins. During therapy, 26 (37.7%) patients had treatment-related adverse events (TRAEs) of any grade. The most common TRAEs were rash (12/69, 17.4%), mouth ulcer (6/69, 8.7%), diarrhea (5/69, 7.2%) and transaminase elevation (4/69, 5.8%). Only 2 (2.9%) patients had TRAEs of grade 3 or higher and 1(1.4%) of the two discontinued therapy. Conclusions: This is the first study to demonstrate that furmonertinib has good efficacy and a tolerable safety profile as adjuvant therapy in EGFR-mutated NSCLC patients with high-risk pathological factors who underwent radical lung cancer surgery.[Table: see text]
Second chance for cure: Stereotactic ablative radiotherapy in oligometastatic disease.
Authors: Salim Nidal , Tumanova Kristina , Popodko Alexey , Libson Eugene
Published: August 2023

154

Background: Local ablative therapy, such as radiation therapy or surgery plays a key role in treatment of patients with oligometastatic disease. Stereotactic ablative radiotherapy (SABR) comes to the fore as a safe and effective treatment option for patients with a limited number of metastases, even those located in hard-to-reach body sites. Many researchers have suggested that metastatic direct therapy (MDT) could improve long-term progression-free and overall survival (PFS and OS) in patients with 1-5 metastatic lesions. Methods: This was a retrospective, single-arm, observational, evaluation study between July 2015 to February 2022. In our institute 60 patients with controlled primary tumors and 1-5 metastases were treated with SABR.We did not exclude patients who subsequently had intracranial metastases from the analysis (n 3). Prescribed radiation doses ranged from 24 to 60 Gy administered in one to seven fractions. Primary endpoint was PFS and the secondary endpoints were OS and toxicity. Results: The most common primary malignancy types were prostate cancer (n 14), colon (n 10), lung (n 7), breast (n 6), melanoma (n 6) and gynecology malignancies (n 6). Median follow-up was 30 months, ranging from 9 to 79. The 1, 3 and 5-year OS rate was 98,3%, 84,4% and 73,8% respectively, the median time to first progression was 15 (range 2 - 32) months. 36 (60%) patients have no recurrence. Two of three patients with intracranial metastases were alive at the end of the study. There were no grade 3-5 adverse events related to specific treatment of SABR. In our analysis, neither the number of metastases nor the primary site were independent prognostic factors. Conclusions: In our retrospective analysis, SABR was associated with high levels of PFS and OS in patients with oligometastatic disease. The limitations of our study were lacking high-level evidence, randomized studies to compare SABR and palliative standard-of-care are mandatory.
Hypoxia-related genes and PD-L1 inhibitor resistance in hepatocellular carcinoma.
Authors: Mohan Huang , Chan Wing Chi
Published: August 2023

158

Background: PD-L1 inhibitor in combination with anti-angiogenic drug has become the first-line treatment strategy for unresectable hepatocellular carcinoma (HCC). However, PD-L1 inhibitor resistance remains an essential issue in treating HCC. It has been proved in the tumor microenvironment that hypoxia-induced increase in PD-L1 expression makes a significant impact on drug resistance. Methods: Two public datasets of gene expression profiles (GSE 14520 and GSE 41666) from the Gene Expression Omnibus (GEO) database were analyzed using bioinformatics: (1) HCC tumor versus adjacent normal tissue (N = 214) and (2) normoxia versus anoxia of HepG2 cells (N = 6). HCC-signature and hypoxia-related genes were identified as the differentially expressed genes (DEGs). Gene set enrichment analyses (GSEA) were performed on the DEGs. Furthermore, multiple regression analysis on the TCGA-LIHC dataset (N=371) and construction of the protein-protein interaction (PPI) network were performed to investigate potential PD-L1 regulatory genes and hub genes. Results: A total of 52 genes overlapped in HCC-signature and hypoxia-related DEGs. Five genes were identified by GSEA as being concurrently associated with the PD-L1 expression pathways. Multiple regression analysis results showed 14 potential PD-L1 regulatory genes. Ten hub genes were identified in the PPI network. Finally, three genes (DLGAP5, KIF20A, and TPX2) were found in common and may be regulatory genes affecting PD-L1 expression. Conclusions: In conclusion, our study provides new insights into the potential hypoxia-related mechanisms of PD-L1 inhibitor resistance and contributes to exploring new therapeutic strategies for treating unresectable HCC.
Comparison of the initial presentation and long-term outcome of differentiated thyroid carcinoma (DTC) between children and adults: A single institution experience in Taiwan.
Authors: Chang Yen Hsiang
Published: August 2023

163

Background: Differentiated thyroid carcinoma (DTC) in children is known to be more aggressive than in adults. However, there are no cohort studies in Asia that have compared the presentation and long-term outcomes of pediatric DTC with adult DTC. In this study, we aimed to compare the clinical characteristics and outcomes of pediatric DTC with a comparison cohort of adult DTC patients. Methods: We reviewed the data of 46 pediatric DTC patients who underwent total thyroidectomy from January 1991 to December 2021, along with a comparison cohort of 350 adult DTC patients at our hospital. All patients received postoperative radioiodine therapy. Clinicopathologic features and surgical outcomes were analyzed retrospectively, and patients were stratified according to the American Thyroid Association (ATA) risk stratification system. Clinical outcomes were classified according to Dynamic Risk Stratification (DRS) at the end of follow-up. Results: Children with DTC had less common papillary thyroid carcinoma (80 vs. 93%) but had larger primary tumors, more multifocal disease, and more lymph node and distant metastasis at presentation compared to adults. Thus, at initial risk stratification, more children had intermediate-to-high risk (62 vs. 49%) than adults. At the end of follow-up, fewer children were classified as having excellent response (no evidence of disease) than adults (48 vs. 79%). Based on the ATA risk stratification system, a higher percentage of high-risk children had structural-persistent disease (60%) compared to high-risk adults (29%), and a lower percentage of low-risk children became no evidence of disease (76%) compared to low-risk adults (93%). Conclusions: Children with DTC present with more extensive disease and have worse outcomes compared to adults. The ATA initial risk stratification system can predict long-term outcomes in both children and adult DTC patients and guide personalized management and follow-up strategies.
Comparison of treatment discontinuation rates between CDK4/6 inhibitors.
Authors: Kosaka Maya , Tanioka Maki , Nishina Takuya , Takata Kenji , Shoko Otsuka , Tsukioki Takahiro , Takahashi Yuko , Iwatani Tsuguo , Kawasaki Kensuke , Shien Tadahiko
Published: August 2023

2

Background: Abemaciclib and Palbociclib are both CDK4/6 inhibitors and its combination use with hormone drugs prolonged progression-free survival in estrogen receptor positive metastatic breast cancer. Both are not yet directly compared in terms of efficacy, adverse events, and discontinuation rates due to the adverse events. MonarchE trial evaluated adverse events of Abemaciclib as follows; 81.3% had diarrhea and 41.3% had neutropenia, whereas PALOMA-3 trial showed 92% of neutropenia (74% had Grade3 neutropenia) in patients receiving Palbociclib. Of the patients who received CDK4/6 inhibitors in our institutions, we compared the difference of rates in adverse events, and we discussed the importance of acknowledging the risks between the two drugs. Methods: We reviewed a total of 169 patients comprising 53 with Abemaciclib and 116 with Palbociclib from 2017 to 2023 at two institutions as 1st – 3rd line treatments. The year of prescription, treatment line, age and the adverse event which led to treatment discontinuation was analyzed. Results: All patients were women, and the median age was 57 (29-84). The patients who received treatment as 1st, 2nd, 3rd lines were 98, 49, and 22, respectively. Twenty-six patients received cytotoxic chemotherapy before CDK4/6 inhibitors. Patients diagnosed as de-novo stage IV patients were 108, while 61 had recurrent tumors. Treatment discontinuation due to adverse events of CDK4/6 inhibitors were 41 (24%) in which 14 (8.3%) had diarrhea and 20 (12%) had neutropenia. We conducted a multivariate analysis setting the endpoint as any adverse events resulting to treatment discontinuation. Abemaciclib (HR=1.20 p=0.01), the first prescription date (2017-19 vs 2020-23, HR:0.84 p=0.02), and patients with metastatic breast cancer at first diagnosis (HR:1.15 p=0.039) significantly affected adverse events leading to treatment discontinuation. Patients receiving Abemaciclib discontinuedecause of diarrhea more than patients receiving Palbociclib (p<0.001 HR:1.28), whereas patients receiving Palbociclib discontinued treatment because of neutropenia more than patients receiving Abemaciclib (HR=1.13 p=0.036). Conclusions: The type of CDK4/6 inhibitor, de-novo Stage IV or recurrent breast cancer, and the year of treatment initiation led to differences in safety of CDK4/6 inhibitor treatment. Abemaciclib leads to treatment discontinuation more than Palbociclib with the most reported adverse event; diarrhea. In addition, the risk of adverse events differs from the year of treatment initiation, which suggests that management of adverse events is improving over time. Thus, it is crucial to accomplish the management of adverse events when starting Abemaciclib which has higher treatment discontinuation rates due to adverse events.
HYbrid DegRAding Copolymer (HYDRAC): Heterobifunctional polymers for targeted degradation of undruggable proteins.
Authors: Wang Max Mu , Truica Mihai Ioan , Abdulkadir Sarki , Gianneschi Nathan
Published: August 2023

22

Background: Of the ~20,000 proteins encoded by the human genome, only a small subset can be modulated via traditional pharmacological approaches. Included in the list of “undruggable” proteins are ones with pivotal roles in cancer. We aim to expand the arsenal of compounds able to affect the activity of these therapeutically relevant targets by introducing a modular proteomimetic platform technology designed to carry side chains consisting of targeting warheads and recruiters of cellular protein degradation machinery. We term these compounds HYbrid DegRAding Copolymers (HYDRACs) in reference to the multiplexing of different protein binders and degradation inducing sequences. Peptides arranged as HYDRACs exhibit favorable emergent properties including resistance to proteolysis, high levels of cell uptake, and payload-specific activity. Initial studies were conducted focusing on a undruggable transcription factor MYC, which is widely dysregulated in human cancers. Methods: A library of HYDRACs and control polymers were synthesized and were cell-penetrating with antiproliferative effects at sub-micromolar IC50s in a formulation- and MYC-dependent manner across multiple cell lines. Results: HYDRAC treatment resulted in significant decreases in MYC protein levels, which was rescued by proteasome inhibitor MG132 and neddylation inhibitor MLN4924, without changes in mRNA levels. RNAseq showed selective overexpression of MYC pathway genes only in HYDRAC treated cells, confirming on-target effects. Circular dichroism showed strong HYDRAC-MYC interactions, which was supported by the presence of MYC following pull-down of biotin-terminated HYDRACs. Removal of the RRRG degron, scrambling the H1 sequence, and disassociation of the two domains on separate polymers all abrogated MYC-degradation. Mice bearing transplanted Myc-CaP tumors showed delayed tumor growth and reduced tumor cell proliferation following HYDRAC treatment. Mice bearing luminescent Luc-MV-411 tumors treated with Cy5.5-labeled HDRACs showed selective tumor accumulation with detectable fluorescent signal in the tumor up to 72 hours following a single IP administration. The versatility of the platform was demonstrated by substituting the degron for recruiters of three different E3 ligases (VHL, KEAP1, and CRBN), which all maintained ability to degrade MYC. HYDRACs were also able to selectively degrade other protein targets including alpha-syn and RAS. Conclusions: We present a platform technology that addresses challenges inherent to small molecule drug design. HYDRACs have the potential to dramatically alter the drug discovery landscape, allowing for development of modulators of "undruggable" targets. We envision the HYDRAC platform as a generalizable approach to designing degraders of proteins of interest, greatly expanding the therapeutic armamentarium for target protein degradation.
A novel approach for the treatment of advanced hepatocellular carcinoma (HCC).
Authors: Fishman Pnina , Lencioni Riccardo , Stemmer Salomon M , Farbstein Motti , Harpaz Zivit , Bareket-Samish Avital , Silverman Michael H. , Ciurescu Ioana Adriana
Published: August 2023

35

Background: No established treatments for patients with advanced HCC and moderate hepatic dysfunction (Child-Pugh B; CPB) are available. Namodenoson is an A3 adenosine-receptor agonist currently under clinical development as a treatment for advanced HCC with CPB. Methods: Wereport the results of a prospective placebo-controlled randomized (2:1) phase 2 trial investigating namodenoson (25 mg BID) as a second-line therapy in HCC with CPB and focus on a case of a long-term complete response (CR). Results: The study included 78 patients (50 namodenoson-treated, 28 placebo-treated). Namodenoson was well-tolerated and no treatment-related deaths were reported. Median overall survival (OS, the primary endpoint) was not statistically significantly different between the namodenoson and placebo arms (4.1 and 4.3 months, respectively, hazard ratio [HR], 0.82; 95% confidence interval [CI], 0.49‒1.38; p=.46). The difference in 12-month OS was statistically significant (44% vs 18%, respectively, p=.028). Pre-planned analysis of patients with CPB7 (34 namodenoson-treated, 22 placebo-treated) showed nonsignificant improvements in OS and progression-free survival (PFS). OS: 6.9 vs 4.3 months; HR, 0.81; 95% CI: 0.45‒1.43; p=.46. PFS: 3.5 vs 1.9 months; HR, 0.89; 95% CI: 0.51‒1.55; p=.67. Of the patients with ≥1 assessment post-baseline, partial response (PR) was reported in 3 of 34 (8.8%) namodenoson-treated patients and none of the 21 placebo-treated patients. One of 3 the patients who achieved a PR (a 61-year old woman with CPB7, Barcelona Clinic Liver Cancer stage C, and Eastern Cooperative Oncology Group performance status 2) continued treatment with namodenoson under an open-label extension program (treatment is ongoing for >6 years). The patient achieved a CR (determined via CT scans) 4 years after treatment initiation. Furthermore, the patient's baseline ALT and AST levels which were elevated at baseline (68 U/L and 44 U/L, respectively) normalized after 4 weeks, and her serum α-fetoprotein level, which was 47 ng/ml at baseline, declined to normal levels after 20 weeks of treatment and reached 1.3 ng/mL at time of CR. The patient experienced no treatment-related adverse events. At present (6 years from treatment initiation), the response is ongoing as shown by liver function evaluation and imaging studies. Conclusions: Treatment with namodenoson can result in a complete and long-term response. The findings support the clinical development of namodenoson as a treatment for HCC with CPB7. Clinical trial information: NCT02128958 .
Identification of the epigenetic mechanism for colorectal cancer stem cell regulation through the comprehensive microRNA expression profiling of human colorectal cancer tissues.
Authors: Mukohyama Junko , Shinoda Masahiro , Itano Osamu , Kakeji Yoshihiro , Shimono Yohei
Published: August 2023

42

Background: MicroRNAs (miRNAs) are key regulators of cancer stem cell (CSC) functions, including self-renewal and differentiation. However, little is known about miRNAs that regulate such properties in human colorectal cancer. Methods: We compared the expression levels of 754 miRNAs between paired samples of EpCAM+/CD44+ cancer cells (enriched in CSCs) and EpCAM+/CD44neg cancer cells (non-tumorigenic cancer cells (NTCs)) sorted in parallel from human primary colorectal cancers by multiplex semi-quantitative PCR. Results: Comparison of the expression levels of miRNAs in the CSCs and NTCs resulted in the identification of 10 miRNAs upregulated (miR-221, miR-218, miR-500 and miR-195) or downregulated (miR-185, miR-19a, miR-337, miR-10a, miR-21 and miR-24) in CSCs compared to NTCs. Among them, miR-221 was most highly expressed in CSCs; and its expression level was very low in NTCs and normal colon epithelial cells (n=6, p<0.05). Kaplan-Myer estimates and univariate and multivariate analyses showed that higher miR-221 expression was an independent prognosis factor for poorer outcome. Constitutive overexpression of miR-221 enhanced organoid-forming capacity of both conventional colorectal carcinoma cell lines and patient-derived xenografts (PDX) in vitro. Importantly, constitutive downregulation of miR-221 suppressed organoid-forming capacity in vitro and substantially reduced the tumorigenic capacity of CSC populations from PDX lines in vivo. miR-221 directly targeted Quaking (QKI) 5, the most abundant splicing isoform of the human QKI gene, and suppressed its protein level. Knockdown of miR-221 suppressed the clonogenicity of colorectal CSCs in vitro in a QKI-expression dependent manner and ability to form tumors in vivo. Conclusions: Our study reveals a mechanistic link between miR-221 and QKI and highlights their key role in regulating CSC properties in human colorectal cancer and proposes that miR-221 will be a promising marker and a specific therapeutic target for human colorectal CSCs.
i-Biomarker CaDx: A circulating miRNA-based multi-cancer detection tool with explainable AI for breast cancer.
Authors: Floares Alexandru George , Zety Adrian Vasile , Floares Carmen , Calin George , Kreutz Carvalho Eduardo , Radi Khaoula
Published: August 2023

5

Background: Breast cancer is requires early and accurate detection for effective treatment and improved patient outcomes. Current methods, such as mammography and biopsy, have limitations in accuracy, invasiveness, and accessibility. Circulating microRNAs (miRNAs) are promising non-invasive biomarkers for cancer detection, including breast cancer. i-Biomarker CaDx is a technology-agnostic, multi-cancer early detection and diagnosis test, covering 13 cancer types with 99-100% accuracy. This study evaluates i-Biomarker CaDx performance for breast cancer using a publicly available dataset and assesses explainable artificial intelligence (XAI) interpretations. Methods: The dataset (GEO: GSE73002) contains 1,280 breast cancer patients and an equal number of age-matched healthy individuals. Circulating miRNA profiles were measured using microarray technology. Multiple classification algorithms, including an ensemble of decision trees, neural networks, and support vector machines, were employed for feature selection, classification, and XAI. Hyperparameter optimization was performed, and the best classifiers were integrated with weights proportional to their performance into a final model. The performance of i-Biomarker CaDx was evaluated using cross-validation and independent test sets, while the XAI component provided cohort-level and individual-level explanations of the miRNA alterations associated with the diagnostic outcome. Results: i-Biomarker CaDx demonstrated remarkable diagnostic performance in detecting breast cancer, achieving 100% accuracy in the studied dataset. The sensitivity, specificity, positive predictive value, and negative predictive value of i-Biomarker CaDx surpassed those of traditional diagnostic methods such as mammography and biopsy. The ensemble classifier, consisting of decision trees, neural networks, and support vector machines, effectively captured the complex interactions among miRNAs. The XAI component of i-Biomarker CaDx provided valuable insights into the biological relevance of miRNA alterations in breast cancer diagnosis. Cohort-level and individual-level explanations elucidated the miRNA patterns and their associations with breast cancer, enhancing the understanding of the underlying molecular mechanisms. Conclusions: i-Biomarker CaDx, a technology-agnostic multi-cancer early detection and diagnosis tool, has shown exceptional accuracy in breast cancer detection, outperforming traditional diagnostic methods. The integration of explainable artificial intelligence (XAI) facilitates a deeper understanding of miRNA alterations and their associations with breast cancer, contributing to improved interpretation at both the cohort and individual levels. The results of this study demonstrate the potential of i-Biomarker CaDx as a powerful, non-invasive diagnostic tool for breast cancer.
Unraveling the heterogeneity of intercellular crosstalk in intrahepatic and extrahepatic cholangiocarcinoma using single-cell and spatial transcriptome profiling.
Authors: Chen Xingyu , Song Yinghui , Tan Zhiguo , Yang Ranzhiqiang , Mao Xianhai , Peng Chuang , Chen Haotian , Liu Sulai
Published: August 2023

50

Background: Extrahepatic cholangiocarcinoma (ECC) and intrahepatic cholangiocarcinoma (ICC) are two types of malignant tumors that arise from the bile duct epithelium at different locations. Currently, effective treatment options for these tumors are limited, and their microenvironmental differences remain poorly understood. Methods: We collected retrospective clinicopathological data of ECC (N=64) and ICC (N=153) patients from our hospital and integrated public database information to analyze the clinical characteristics, survival, and prognosis of ECC and ICC. We performed single-cell sequencing (scRNA-seq) and spatial transcriptomic techniques on three ECC and three ICC tumor tissues. Verification of scRNA-seq results was done using immunohistochemistry, flow cytometry, and polychromatic immunofluorescence. Additionally, we analyzed RNA and ATAC sequencing data for exhausted CD8+ T cells TEX obtained from ICC and ECC tissue samples. We used InferCNV to infer copy number variations (CNVs) of epithelial cells from ICC and ECC. Results: Our results showed that ECC and ICC differ in their clinical characteristics such as age, gender, bilirubin, CEA, and outcome. ECC had higher copy number amplification variation and exhibited inflammatory signaling and ongoing immunoediting, reflected in higher immune checkpoint expression such as PDCD1, CTLA4, and LAG3, and tumor infiltration with highly differentiated dysfunctional PDCD1+ LAG3+ CXCL13+ exhausted CD8+ T cells (TEX) and precursors of exhausted T (TPEX). RNA velocity analysis revealed different developmental trajectories of T cells in ECC and ICC, predominantly in naive/stem-like and memory T cells. We also identified a trefoil factor 3 (TFF3)-positive epithelial subpopulation in ECC. TFF3+ epithelial cells had closer interaction with CTHRC1+ fibroblasts, which induced more CD8+ T cells to TEX and TPEX. TFF3+ cells exhibited higher CNV scores and higher PD-L1 and PD-L2 expression, and co-localized with TEX and TPEX, providing one explanation for the immunosuppressive environment in ECC and suggesting a potential response to immune checkpoint therapy (ICT). Conclusions: Our study provides a comprehensive characterization of the cellular heterogeneity landscape of ECC and ICC, which can guide the optimization of personalized cancer immunotherapy. We identified potential biomarkers and targets for immunotherapies, and our findings shed light on the underlying mechanisms of tumor immune evasion and provide a foundation for further research in this area.
Single cell profiling of tumour biopsies and heterogeneity in response of dedifferentiated melanoma.
Authors: Lim Esther , Lin Yingxin , Pedersen Bernadette , Stewart Ashleigh , Scolyer Richard A. , Long Georgina V. , Yang Jean , Rizos Helen
Published: August 2023

64

Background: Patients with advanced BRAFV600-mutant melanoma can be treated with immune checkpoint inhibitors or combination BRAF/MEK inhibitors. The sequence of these therapies can influence response to second-line treatment. Patients who progress on first-line PD1 blockade have poorer outcomes on BRAF/MEK inhibitors compared to patients who are treatment-naïve. Thus, defining the molecular features that are modified by therapy and that influence subsequent treatment responses require thorough characterization. Methods: In this study, tumor biopsies from five BRAF-mutant melanoma patients progressing on anti-PD1 (PROG) and five BRAF-mutant treatment-naïve (NAÏVE) patients were dissociated and treated ex vivo with BRAF/MEK inhibitors (dabrafenib+trametinib (DT)) before flow cytometry profiling and single cell transcriptome sequencing. Results: DT treatment did not alter tumor or immune cell proportions but effectively inhibited MAPK signaling by decreasing phosphorylated S6 levels in tumor cells within 8/10 melanoma samples. Single cell RNA-sequencing of the tumor dissociates identified four distinct melanoma differentiation states (e.g. melanocytic, transitory, neural crest-like and undifferentiated). The proportions of these cell states differed between tumor specimens, and were not affected by DT treatment. The neural crest-like and undifferentiated melanoma cells were enriched in PROG compared to NAIVE tumor samples and all differentiation states responded, albeit variably. For instance, MAPK inhibition after DT treatment was less pronounced in undifferentiated melanoma cells. Importantly, within the undifferentiated cell state, we identified a proportion of melanoma cells capable of responding to DT, and comparison of responding vs non-responding undifferentiated cells showed enrichment of pro-inflammatory IL-6 and TNFα signaling in the non-responding population. Elevated IL-6 and TNFα pathway activity correlated with decreased potency to MAPK inhibition, indicating that increased IL-6 and TNFα signaling in undifferentiated melanoma may contribute to resistance to MAPK inhibition. Conclusions: Ex vivo treatment of tumor dissociates coupled with single cell transcriptome sequencing identified cell state-specific responses to MAPK inhibition. Resistance to MAPK inhibition may be driven, in part, by tumor-intrinsic pro-inflammatory IL-6 and TNFα signalling.
Her-2 promoting tumor progression and cisplatin resistance as a novel therapeutic target in penile squamous cell carcinoma.
Authors: Tan Xingliang , Yao Kai
Published: August 2023

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Background: Cisplatin-based chemotherapy has been the first choice for advanced penile squamous cell carcinoma (PSCC) in the last decade, but remains challenging owing the unsatisfied response rates, frustrating systemic toxicity and chemoresistance leading to poor prognosis. There is no standard second-line therapy for advanced PSCC. Antibody-drug conjugates (ADC) were novel targeted and low-toxicity agents, of which Her-2 targeted ADCs widely used and have greatly improved clinical outcomes in breast, gastric and bladder cancers. We aimed to explore the expression pattern, clinical significance, oncogenic roles of Her-2 and the therapeutic potentiality of Her-2 target ADCs in PSCC. Methods: The immunohistochemical expression of Her-2 was detected in the largest single center cohort of 367 PSCC patients up to date. PSCC cell lines, cisplatin-resistant cell lines and animal models were established to investigated the biological roles of Her-2 in PSCC progression and cisplatin chemoresistance. Disitamab vedotin (RC48), an Her-2 targeted ADC drug, was used to evaluate the efficiency in PSCC. Results: Her-2 was an unfavourable prognostic indicator with around 47.7% immunohistochemical expression rate (1+, 23.2%; 2+, 18.0%; 3+, 6.5%) in PSCC. Her-2 positive immunohistochemical expression (2+/3+) was associated with advanced pT, pN stages, metastasis and poor overall survival (positive vs. negative, HR (95%Cl): 3.133 (2.120-4.631), p=0.000). Cellular and animal experiments confirmed that Her-2 promoted tumor progression, inhibited cisplatin-induced cell apoptosis via Akt pathways and increased cisplatin resistance in PSCC. Meanwhile, cisplatin-resistant PSCC cells up-regulated Her-2 expression. More importantly, Her-2 targeted ADCs Disitamab Vedotin displayed remarkable anti-tumor activities both in Her2-positive and cisplatin-resistant PSCC tumors. Conclusions: Our study suggested that Her-2 was an available therapeutic biomarker in PSCC. Targeting Her-2 ADCs might be a prospective option to improve clinical outcomes in high-risk Her-2 positive advanced PSCC patients and provided precious opportunity for cisplatin-based chemoresistance patients.
Risk factors and predictors of tumor recurrence among breast cancer patients in a tertiary government hospital in the Philippines.
Authors: Maaño Omar Paningbatan , Soliman Mary Claire V.
Published: August 2023

7

Background: This study aims to identify the risk factors and predictors of tumor recurrence of breast cancer patients using the local data available in the Breast Care Center in East Avenue Medical Center, Quezon City, Philippines. Methods: A chart review of 529 charts was done at the Breast Care Center between 2011–2017, and 474 charts were included. Demographics and Clinicopathologic features were gathered from the chart. Cox-proportional hazard regression was used to compute hazard ratios [95% confidence interval] of the different variables to compare the recurrence rates. Results: Breast cancer subtypes are as follows: 246 (52%) patients were HR+/Her2-, 52 (11%) were HR-/Her2+, 81 (17%) were HR+/Her2+, and 95 (20%) were HR-/Her2-. The mean age at diagnosis was 50 years old. The median BMI was 25 kg/m2. Most have hypertension (HTN) (21.7%) and diabetes mellitus (DM) (7%). The most common histology was invasive carcinoma NOS (92.6%), and 42.5% had a lymphovascular invasion. Only 6.7% received neoadjuvant systemic therapy, the rest had adjuvant systemic therapy. Mostly underwent modified radical mastectomy (92.8%). The most common cytotoxic agent used were cyclophosphamide (96.14%), doxorubicin (87.13%), docetaxel (53.37%), and 5-fluorouracil (37.87%). In the her2+ subtype, only 19.54% received trastuzumab. Among breast cancer subtypes, HR-/Her2+ has the highest tumor recurrence (15.38%), and HR+/Her2-, HR+/Her2+, and HR-/Her2- had 8.54%, 7.41%, and 11.96% had tumor recurrence, respectively. The median time to recurrence was 47, 24, 39.5, and 47 months with HR+/Her2-, HR-/Her2+, HR+/Her2+, and HR-/Her2-, respectively. The most common site of recurrence were bone (39%) and local recurrence (32.37%). Contralateral breast recurrence occurred in 2 patients with HR-/Her2+. In univariate analysis, HTN (HR 0.34 [0.13, 0.85], p=0.022), DM (HR 2.55 [1.19, 5.48], p=0.016), stage IIIA (HR 3.78 [1.04, 13.73], p=0.044), IIIB (HR 9.11 [2.46, 33.68] p=0.001), IIIC (HR 12.59 [3.50, 45.22], p<0.001), positive lymph nodes (HR 1.07 [1.02, 1.12] p=0.002), and patients who received doxorubicin (HR 0.44[0.23, 0.86], p=0.015) were statistically significant for tumor recurrence. In multivariate analysis, HTN (HR 0.32 [0.11, 0.94], p=0.038) and doxorubicin (HR 0.30 [0.13, 0.70], p=0.005) have the strongest effect for decreasing recurrence rate and DM (HR 7.45 [2.85, 19.46] p<0.001) and locally advanced stage (IIIB) (HR 10.97 [2.51, 48.06], p=0.001) were associated with greater risk of recurrence. Conclusions: In this study, we determined the risk factors and predictors of breast cancer tumor recurrence in a tertiary government hospital in a third-world country like the Philippines. Certain co-morbidities (HTN and DM), locally advanced stage at diagnosis, and systemic treatment received (doxorubicin) significantly affected tumor recurrences with breast cancer in our setting.
Using sacituzumab govitecan for metastatic urothelial carcinoma after prior treatment with enfortumab vedotin: A single institution experience.
Authors: Maroli Kimberly , Wee Christopher Eing , Nizam Amanda , Ornstein Moshe Chaim , Gilligan Timothy D. , Martin Allison , Bonham Amanda , Lynn David , Vyas Neil , Gupta Shilpa
Published: August 2023

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Background: The Trop-2-directed antibody drug conjugate sacituzumab govitecan (SG) has demonstrated efficacy in metastatic urothelial cancer (mUC) previously treated with checkpoint inhibitors +/- prior chemotherapy based on the results from the TROPHY U-01 study. However, only 10/118 and 7/38 patients in the reported Cohorts 1 and 2 respectively had received prior enfortumab vedotin (EV). Given that EV is preferred prior to SG given proven overall survival (OS) benefit vs chemotherapy, we analyzed our institution’s experience to determine real-world outcomes when SG is used after prior EV treatment. Methods: We identified patients who had received EV for mUC at our institution and identified a subset of patients who had received SG after prior treatment with EV. Patient characteristics (e.g. age, sex, primary site of disease) were noted, as well as the number of lines of prior systemic therapy for mUC. Best response to SG was recorded, as well as time to progression, reason for discontinuation, and time to death. Progression free survival (PFS) was calculated from time of SG treatment initiation until progression or death, and overall survival (OS) was calculated from time of SG treatment initiation until death. Results: Ten patients were identified having been treated with SG for mUC after prior EV treatment. Seven out of ten were male and the median age at time of initial EV treatment was 68 years. Half of the patients had the primary upper tract urothelial carcinoma. There were no responses to treatment with SG; the best response to SG treatment was stable disease (SD) in 3/10 patients, progressive disease in 4/10 patients, and three patients’ responses were not assessed due to death or transition to hospice at time of last follow-up. The median PFS was 2.5 months and median OS was 3.2 months. Only two patients survived more than six months (7.1 months and 10.8 months). Conclusions: In this small retrospective series at a single institution, real-world outcomes (response to treatment, PFS, OS) using SG for mUC after prior EV treatment appear to be worse than the reported outcomes in TROPHY-U01 trial. The lack of prior EV treatment in the majority of the SG clinical trial patients could contribute to these disparate results.
Deep learning based quantitative models to distinguish tumoral and stromal tumor-infiltrating lymphocytes and the prognosis patients with high-grade serous ovarian cancer.
Authors: Hamada Kohei , Murakami Ryusuke , Ueda Akihiko , Taki Mana , Yamanoi Koji , Yamaguchi Ken , Hamanishi Junzo , Mandai Masaki
Published: August 2023

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Background: The better prognostic value of tumor-infiltrating lymphocytes (TILs) has been reported in high-grade serous ovarian cancer (HGSOC). However, the TILs evaluation of previous reports largely depends on manual quantification based on a visual assessment within limited section of pathological slides or tissue microarrays, which is subject to inter-observer variability. Since tumor immune microenvironment (TIME) of HGSOC is highly heterogeneous, there is a need to quantify TILs in large areas of pathological slides in an objective and time-saving manner. Here we present an AI-powered quantification method to distinguish tumoral TILs and stromal TILs using hematoxylin and eosin (H&E)-stained whole-slide images (WSIs) of HGSOC. Methods: 161 H&E stained WSIs of HGSOC patients treated at our hospital between 1998 and 2021 were prepared, and by using these WSIs we generated two AI-powered models. Initially, a TILs detection model (TIL-model) was created using a publicly available dataset of 86104 pan-cancer TILs images of 100 × 100 pixels at magnification of x200 (Saltz et al., 2018, Cell Reports). Secondary, a tumor and stroma segmentation model (T/S seg-model) was developed using 136962 patches of 224 × 224 pixels at magnification of x50 which were manually annotated with tumoral regions and stromal regions in 102 H&E stained WSIs of HGSOC from The Cancer Genome Atlas (TCGA). Deep convolutional neural networks of NASNet-Large for the TIL-model and TransUNet for the T/S seg-model were used. TILs density score (number of TILs positive tiles per tumor area) of each sample was calculated, and association between TILs density score and the prognosis was investigated. Results: The established models achieved high accuracy: the TIL-model showed an area under the curve (AUC) value of 0.896, and the T/S seg-model showed an intersection over union value of 0.967. By combining the two models, we successfully distinguished tumoral TILs and stromal TILs. Using our dataset of 161 WSIs, we split the patients by TILs density score at top quartile into high tumoral TILs subgroup (n=40) and low tumoral TILs subgroup (n=121). High tumoral TILs subgroup showed significantly longer overall survival (OS) and progression free survival (PFS) (OS: median 70 months vs not reached, p < 0.005. PFS: median 34 months vs 21 months, p = 0.02). Conclusions: We developed deep learning based pathological AI models to detect TILs and tumors using H&E stained WSIs, which achieved to quantify tumoral and stromal TILs. The objective assessment of TILs using these models will lead to the development of clinically applicable tools for the evaluation of the immune status in HGSOC.