bluuebunny/crossref_metadata_2025 huggingface dataset:
The rationale of reprogramming TREM2+ macrophages for overcoming the resistance to immune checkpoint inhibitors in pancreatic adenocarcinoma.
Authors: Ravichandran Harshitha , Kung Heng-Chung , Zhang Rui , Chen Kai , Fu Juan , Zheng Lei
Published: June 2024

e14649

Background: Pancreatic ductal adenocarcinoma (PDAC) resists to immune checkpoint inhibitors as single agents. TAMs expressing the triggering receptor expressed on myeloid cells (TREM2) protein were found to be a dominant macrophage population in PDAC; targeting TREM2 has been found to enhance the efficacy of immune checkpoint inhibitor-based therapies in preclinical studies of other malignancies. Here, we studied the role of targeting TREM2 in PDAC by testing the combination of non-depleting anti-TREM2 antibody and anti-PD-1 antibody in a mouse model of PDAC. We also examined the relationship between TREM2 expression and effector T cells in surgically resected human PDAC collected from a previous clinical trial treated with the neoadjuvant combination therapy of nivolumab (anti-PD-1 antibody), urelumab (anti-CD137 agonist antibody), and the pancreatic cancer GVAX vaccine. Methods: C57BL/6 mice were orthotopically implanted with KPC tumors. Anti-TREM2 was injected intraperitoneally (i.p.) once a week and anti-PD-1 was injected i.p. twice a week. Tumor volume was measured by ultrasound. TREM2 was stained on slides which were previously obtained from surgically resected PDACs following the neoadjuvant combination treatment of nivolumab, urelumab, and GVAX and stained for multiple immune cell markers with a multiplex staining-striping immunohistochemistry (IHC) technique. Results: Tumor growth post-tumor implantation was found to be significantly decreased by the anti-PD-1 and anti-TREM2 combination compared with anti-PD-1 alone, anti-TREM2 alone, or control treatment (p < 0.0001). Survival of tumor-bearing mice was significantly prolonged by the anti-PD-1 and anti-TREM2 combination when compared to anti-PD-1(p < 0.05), anti-TREM2(p < 0.05), and control (p < 0.01). Multiplex IHC analysis of human PDACs following neoadjuvant immunotherapy showed a higher density of granzyme B+CD8+ effector T cells correlated with higher density of TREM2+ cells (p = 0.0357). Conclusions: Our study demonstrates the anti-tumor efficacy of anti-TREM2 antibody in combination with an immune checkpoint inhibitor in mouse PDAC. However, the analysis of human PDAC following neoadjuvant immunotherapy suggests that TREM2 on TAMs may be activated following the activation of effector T cells. Taken together, our results suggest that TREM2+ TAMs should be reprogrammed instead of being depleted in the design of anti-TREM2-based therapy for PDAC.
Advancing drug development by leveraging microdissection for proteogenomic analysis of histological sections.
Authors: Johann Donald Joseph , Shin Ikjae Jae , Tangrea Michael , Emmert-Buck Michael
Published: June 2024

e14660

Background: Combining laser capture microdissection (LCM) with proteogenomics in cancer research offers a targeted way to way to explore the intricate interactions between the tumor microenvironment and tumor cells proper. For immuno-oncology (IO) research, this is especially important, since improvements in the predictability of patient response to IO-based drugs is greatly needed. An improved understanding of the spatial relationships involving tumor cells, stromal cells, blood supply and immune cell interactions may help to inform drug development and improve the indicators of success for IO-based drug regimens. Methods: From fresh frozen tissue, LCM is used to isolate and obtain distinct histological cell types. Nucleic acids and proteins are extracted from captured cells for in-depth multi-modality molecular profiling assays. This includes multiple genomic modalities and a customized solution-based mass spectrometry proteomics approach for samples or specimens with a limited amount of tissue. Results: Optimizing the analysis of minute tissue quantities from LCM captured cells is challenging. Following the isolation of nucleic acids, DNA sequencing can be performed for the discovery and analysis of actionable mutations, copy number variation, and methylation profiles. RNA-seq may be performed for gene expression. Regarding genomic-based studies, commercial kits are often utilized for these steps due to their general availability, rapid innovation, and the robust performance of these products. However, this kit-based adaptive approach is not true for proteomics. There remains a need for highly sensitive proteomic methods targeting small-sized samples. Our novel proteogenomics approach is focused around a customized enhanced liquid chromatography mass spectrometry (LC-MS) analysis of micro-scale and/or nano-scale tissue sections. This is achieved with a silver-stained one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) approach developed for LC-MS analysis of fresh-frozen tissue specimens obtained via LCM. This includes an in-gel digestion method adjusted and specifically designed to maximize the proteome coverage from amount-limited LCM samples to better facilitate in-depth molecular profiling. Conclusions: A proteogenomic approach is reported that is utilizing amount limited tissue quantities from microdissected fresh frozen tissue. The methodology may also be applicable to other types of specimens having limited nucleic acids, protein quantity, and/or sample volume.
Melanoma tumor microbiome as a predictor of anti-PD-1 and anti-CTLA-4 immunotherapy response.
Authors: Uzelac Matthew , Li Wei Tse , Li Yuxiang , Ongkeko Weg M.
Published: June 2024

e14666

Background: In 2023, it is estimated that 97,610 melanoma diagnoses were made in the United States, resulting in 7,990 deaths. For high-risk melanomas, immune checkpoint inhibitors are often administered as an adjuvant to surgery, though they yield variable response rates. The gut microbiome’s association to these treatments’ success has been demonstrated, largely through fecal transplant and probiotic trials. In this study, we seek to determine whether tissue-resident microbes in melanoma tumors could be used to predict immunotherapy response. Methods: Sequencing data of 289 melanoma tumor tissue samples was downloaded across three distinct studies. Intratumor microbial species abundances were derived from RNA or DNA sequencing data through direct alignment to microbial reference databases. Gene expression profiling was also performed. Microbial abundance values were used to construct models predicting patients’ responses to check-point blockade immunotherapies, which were then validated on an external dataset. Results: 12 and 14 microbial species were differentially abundant in anti-PD-1 responders and anti-CTLA-4 responders, respectively. We identified significant correlations of these species to genes of the PD-1 and CTLA-4 signaling pathways. Among others, greater abundance of Corynebacterium kroppenstedtii and Streptococcus gordonii correlated to decreased expression of PD1 and CTLA-4. With the abundances of these taxa and the expressions of 37 genes known to be implicated in immunotherapy response, we developed two classifiers to predict a patient’s response to anti-PD-1 and anti-CTLA-4 therapies. When tested externally, these models successfully predicted a majority of responders and non-responders with sensitivities of 80.0% and 60.0% and specificities of 95.2% and 95.8%, respectively. Conclusions: Our study revealed that checkpoint blockade response for melanomas could be predicted with microbe abundances. Our analysis demonstrates the potentially promising clinical utility of intratumor microbiota in cancer treatment. Given their location in the tumor microenvironment and their ability to influence immune processes, intratumor microbes could play a critical role in modulating the therapeutic effects of immune checkpoint blockades. We hope that our results could also be applied to other solid tissue cancers, where a significantly smaller fraction of patients respond to immunotherapy. [Table: see text]
First-in-human study of ZGGS15, a dual specific antibody targeting LAG-3 and TIGIT, as monotherapy in patients with advanced solid tumors.
Authors: Zhu Ji , Wang Zhen , Chai Xiaoli , Wu Lihua , Qu Song , Liu Linlin , Liu Yanyan , Sun Yan
Published: June 2024

e14678

Background: ZGGS15 is a recombinant humanized bispecific antibody composed of a monoclonal antibody against LAG-3 and TIGIT. It can specifically bind LAG-3 and TIGIT. It reverses LAG-3 and TIGIT immune suppression to T and NK cells, relieves suppression of Treg to T and NK cells, promotes the activation and proliferation of T and NK cells, and cytokines release. ZGGS15 restores T and NK cell functions to kill tumor cells. In non-clinical studies, we found that ZGGS15 had significant anti-tumor effects and synergistic anti-tumor effects with Anti-PD-1 antibody. Therefore, we conducted a Phase 1 clinical study to assess the tolerability, safety, and preliminary efficacy of this drug as monotherapy in patients with advanced solid tumors. Methods: During the dose escalation stage, an accelerated titration design (ATD) in the first dose (1 subject if ≤Grade 2 treatment-related adverse event occurred) and the standard "3+3" design in the rest doses were used. Subjects were patients with advanced solid tumors who failed to the available standard treatments. The dose groups were set from 0.3 to 30 (0.3, 1, 3, 10, 20, 30) mg/kg, intravenous infusion, once every 3 weeks. The first 21-day period was defined as the dose-limiting toxicity (DLT) observation period. Subjects could continue to receive ZGGS15 until treatment discontinuation criteria were met. Tumor response was assessed by RECIST1.1 and iRECIST. Pharmacokinetics (PK) and receptor occupancy (RO) profiles were also assessed in this study. Results: A total of 12 patients (4 males and 8 females) completed the DLT observation in the first four dose groups (0.3 to 10 mg), with a median age of 57 years. Eight (66.7%) of them received at least 3 lines of antineoplastic therapies, and five (41.7%) have been exposed to PD-1 or PD-L1 inhibitors. There was no DLT event observed. Thirty-eight treatment-related adverse events (TRAEs) occurred. Up to now, TRAEs were all grade 1 ~ 2. The most frequent TRAEs were proteinuria (33.3%, 4/12), hypokalemia (25%, 3/12), hypoalbuminemia (25%, 3/12), and hypertriglyceridemia (25%, 3/12). The Cmax and AUC increased from 0.3-1 mg/kg approximately in dose proportion. Nearly complete and sustained RO of T and NK cells in PBMCs was observed at 1 mg/kg dose of ZGGS15 for at least 3 weeks. Conclusions: The first four dose levels of ZGGS15 were very well tolerated. A good PK profile, and a long-lasting RO were observed in this escalation study. It is anticipated that this product could provide an additional benefit only when use in combination with an anti-PD-1 antibody. Clinical trial information: NCT05864573 .
Bronchoalveolar lavage fluid analysis in patients with checkpoint inhibitor pneumonitis.
Authors: Chen Ruxuan , Shi Yujie , Huang Hui , Wang Mengzhao , Xu Yan
Published: June 2024

e14683

Background: Checkpoint inhibitor pneumonitis (CIP) is a relatively uncommon but potentially life-threatening immune-related adverse event (irAE). Lung biopsies have not been commonly performed for CIP patients. Bronchoalveolar lavage fluid (BALF) analysis is a useful diagnostic approach for interstitial lung disease. However, BALF features were inconsistent across different studies. Methods: We retrospectively reviewed the medical records of 154 patients with pathologically confirmed malignancies and suffering from CIPs between July 2018 and December 2022. Patients who had bronchoalveolar lavage (BAL) data available were enrolled in our study. Patient clinical, laboratory, radiological and follow-up data were reviewed and analyzed. In our study, patients with grade 1 or grade 2 CIPs were defined as low-grade CIPs, and patients with grade 3, grade 4 or grade 5 CIPs were defined as high-grade CIPs. Results: The BALF differential cell count and lymphocyte subset analysis were performed for 42 CIP patients. There were 32 males (76.2%). The mean age at diagnosis of CIP was 62.0 ± 10.4 (range: 31-78) years. The median time to onset of CIP was 98.5 days after the start of immunotherapy. Most of the patients were diagnosed with lung cancer (36 patients, 85.7%). There were 3 patients with esophageal squamous cell cancer, 1 patient with hepatic cell cancer, 1 patient with gastric signet ring cell cancer and 1 patient with pericardial malignant mesothelioma. There were 18 patients (42.9%) with low-grade CIPs and 24 patients (57.1%) with high-grade CIPs. The mean lymphocyte percentage was 36.7 ± 22.5%. There were 34 (81%) CIP patients with a lymphocytic cellular pattern. The median ratio of CD3+CD4+/CD3+CD8+ lymphocytes was 0.5 (0.3, 1.0). The ratio was less than 1.0 for 31 CIP patients (73.8%). However, there was no significant difference in the BALF features between patients with low-grade CIPs and those with high-grade CIPs. Conclusions: The CD3+CD8+ lymphocytosis pattern was the main inflammatory profile in the BALF of CIP patients in this cohort. Targeting CD3+CD8+ lymphocytes might be a treatment option for CIPs.
Immune related adverse events: Timing, identification patterns, and impact on survival.
Authors: Deviley Jake Anthony , Hayashi-Tanner Yacki , Krause Joshua , Arenz Andrea , Frankki Susan M , Marinier David E.
Published: June 2024

e14705

Background: Increased use of immune checkpoint inhibitors (ICI) has resulted in a rise in immune related adverse events (irAEs), which many non-oncology clinicians may underrecognize. Controversy exists surrounding whether irAE presence has an impact on survival. Our primary goal was to evaluate the timing between detection to diagnosis of irAE, and determine the department responsible for making the irAE diagnosis. We also assess the incidence of single vs multisystem irAEs, impact of irAE on survival, and assess whether standard irAEs (s-irAEs) those diagnosed within 90 days of initial ICI administration vs delayed irAE (d-irAEs) those diagnosed >90 days impacts survival. Methods: We performed an IRB approved retrospective study of patients 18 years and older who received ICIs for a malignancy at Gundersen Health System between January 1, 2011 and December 31, 2021. Statistical analysis included Chi-square, Fisher’s Exact, and Wilcoxon rank sum tests and Kaplan-Meier survival analysis. All analysis was completed in R version 4.2.3 with a p-value <0.05 considered significant. Results: Our study included 664 patients who were 99.6% white, 60% male, and an average age of 67 years. Median Charlson Comorbidity Index was 10. There were 245 (36.8%) patients with at least 1 irAE, and 62 (9.3%) with a multisystem irAE. While 28 (11%) irAEs were initially detected by non-oncology providers, only 10 (4.1%) irAEs were diagnosed by a non-oncology department. Median time (days) from symptoms to diagnosis was 3 for those diagnosed in non-oncology departments and 0 for those diagnosed in oncology (p<0.01). A total of 142 (58.0%) s-irAEs were detected with grades 1-2-3-4-5 accounting for 21.1%-54.9%-19.7%-1.4%-2.8%. There were 103 (42.0%) d-irAEs with grades 1-2-3-4-5 accounting for 23.3%-53.4%-17.4%-5.8%-0%. Of the d-irAEs, 17 (16.5%) occurred >365 days from initiation of ICI. The 3 most common irAE in this patient population included thyroiditis (34%), dermatitis (21%), and pneumonitis (12%). Our data shows a significant survival difference in patients with an irAE compared to those without (p<.01). No significant survival difference was noted between patients who had s-irAE vs d-irAE (p=.069). Conclusions: Our results show that irAEs may be underrecognized and often not formally diagnosed by non-oncology providers. However, median time to diagnosis is short within this context. This may be due to non-oncology providers feeling uncomfortable making the final decision, which emphasizes the importance for oncologists to finalize irAE diagnoses. We did see a positive impact on survival with irAE presence. While oncologists often clinically expect irAEs to surface within the first 6-10 weeks of treatment, our study demonstrates that 42% of irAEs occur after 12 weeks from ICI initiation. There is no difference in survival between s-irAEs vs d-irAEs, and overall grade distribution between these two groups appears similar.
Increased risk of hyperglycemia in patients with advanced urothelial cancer treated with enfortumab vedotin: A systematic review and meta-analysis.
Authors: Cheng Ryan , Boparai Montek , Wu Shenhong
Published: June 2024

e15001

Background: Enfortumab vedotin (EV), an antibody-drug conjugate targeting anti-nectin-4, which is highly expressed in urothelial cancers, has emerged as an effective therapy in the management of previously treated advanced urothelial cancer. urothelial cancer. Severe hyperglycemia has been recognized as an unexpectedly adverse effect, and can lead to discontinuation of EV or fatal event. To assess the overall risk of hyperglycemia associated with the use of EV, a systematic review and meta-analysis of published clinical trials was performed. Methods: A comprehensive review through 2023 of PubMed and American Society of Clinical Oncology conferences was performed to identify relevant clinical trials and abstracts. Eligible studies were clinical trials of patients with urothelial cancer receiving EV at the standard dose with data on hyperglycemia available. Incidence rates, relative risks, and 95% confidence intervals were calculated using random-effects or fixed-effects models. Results: Our search culminated in the inclusion of 1,150 patients from four trials, with 859 of them receiving EV. The summary incidences for all-grade and high-grade hyperglycemia were 9.9% (95% CI: 7.8-12.5%) and 6.0% (95% CI: 4.6-7.8%) respectively. Patients treated with EV had an increased risk of developing hyperglycemia in comparison with chemotherapy with an RR of 19.66 (95% CI: 2.66-145.55, P=0.004) for all-grade and 26.54 (95% CI: 1.59-444.48, P=0.023). In comparison with single-agent EV, its combination with pembrolizumab did not significantly increase the risk of all-grade hyperglycemia (P=0.148). Conclusions: EV is associated with a significantly increased risk of all-grade and high-grade hyperglycemia.
BC3195, a novel ADC targeting CDH3: Preliminary results of a first-in-human phase I study in patients with advanced solid malignancies.
Authors: Chen Hua-Jun , Wang Sheng , Su Rong , Li Xu-Feng , Tang En-Tzu , Wang Yi-Wei , Hei Yong-Jiang , Wu Yi-Long
Published: June 2024

e15008

Background: Cadherin-3 (CDH3), a calcium-dependent cell-cell adhesion glycoprotein, is overexpressed on lung, breast, ovarian, colorectal, pancreatic, head and neck and other malignancies, but with negligible expression on nonmalignant tissues, and associated with cancer aggressiveness, invasiveness, and poor prognosis. BC3195 is known as the only antibody drug conjugate (ADC) in clinical stage, targeting CDH3 with payload of monomethyl auristatin E (MMAE). Methods: A phase I, open-label, first in human study to evaluate the safety, tolerability, PK, and preliminary antitumor activity of BC3195 is being performed in subjects with advanced solid malignancies. Disease assessments are performed every 6 weeks using RECIST v1.1. BC3195 is administered as 1 hour (h) IV infusion every 3 weeks. An evaluation of seven dose levels is planned: 0.3, 0.6, 1.2, 2.4, 3.0 and 3.6 mg/kg with a BOIN design guiding dose escalation. Results: Up to January 1st, 2024, nine patients (median age, 59; male, 56%) have been enrolled and treated with BC3195, with 3 patients each in the 0.3, 0.6 and 1.2 mg/kg dose cohorts. No DLT was observed across all the 3 dose cohorts. Treatment emergent adverse events (TEAEs) occurring in ≥3 patients included: aspartate aminotransferase increased (5/9, 55.6%), conjugated bilirubin increased (5/9, 55.6%), triglycerides increased (4/9, 44%), uric acid increased (3/9, 33.3%), heart rate increased (3/9, 33%), and hypoalbuminemia (5/9, 55.6%), cough (4/9, 44.4%), hyponatraemia (4/9, 44.4%), anemia (4/9, 44.4%), asthenia (3/9, 33.3%), rash (3/9, 33.3%), vomiting (3/9, 33.3%) and back pain (3/9, 33.3%). Two subjects experienced ≥ Grade 3 TEAEs unrelated to study drug. Six subjects were evaluable for tumor assessment, and 3 subjects (3/6, 50%) show stable disease (SD) as the best response (2 subjects with target lesion reduction). PK results demonstrated that exposure for the ADC and total antibody (TA) increased with dose, the relationship between AUC0-last and dose was greater than dose-proportional, while Cmax was linear. Free MMAE was highest at the 1.2 mg/kg dose level. Median Tmax values for ADC and TA were 1 h (end of infusion), and median Tmax for free MMAE was 25-169 h. In addition, elimination t1/2 values averaged 27-48 h, 31-68 h and 63-76 h for the ADC, TA, and MMAE, respectively. Conclusions: BC3195 exhibited favorable safety profile and PK characteristics up to 1.2mg/kg. Given BC31095’s safety and PK behavior, as well as preliminary activity at the 0.6 mg/kg dose level, patient enrollment at higher dose levels is ongoing. Clinical trial information: NCT05957471 .
Type III collagen pro-peptides (PRO-C3) as a proposed prognostic biomarker of risk of survival outcomes in clinical trials of patients with solid tumors.
Authors: Willumsen Nicholas , Nissen Neel Ingemann , Karsdal Morten Asser
Published: June 2024

e15050

Background: Tumor fibrosis is essential for defining outcome of patients with various solid tumors. The major pathological signature of tumor fibrosis is a fibrous connective tissue formed by proliferation and activation of fibroblasts and increased synthesis and deposition of fibrillar collagens (type III collagen) leading to immune exclusion and high interstitial pressure in the tumor microenvironment. Recently, the USA FDA issued a Letter of Support to encourage the further study and use of the biomarker PRO-C3 (type III collagen pro-peptides) as a proposed prognostic biomarker of risk of survival outcomes in clinical trials of patients with solid tumors ( https://www.fda.gov/media/169332/download ). Here we summarize the published data that supports the use of PRO-C3 as a prognostic biomarker that reflects fibrotic activity in the TME. Methods: A PubMed search was used to find studies with PRO-C3, cancer and survival outcome. The hazard ratios (HR) with 95% confidence intervals (95%CI) calculated from univariate cox regression analyses for overall survival (OS) outcomes were collected for where patients were dichotomized based on their pre-treatment PRO-C3 levels and results summarized across indications including breast cancer, pancreas cancer, colorectal cancer, hepatocellular carcinoma, biliary tract cancer and malignant melanoma. Results: Nine studies were retrieved. Table 1 shows the studies with HR and 95%CI for OS after dividing patients into high and low PRO-C3. Various cutoffs were used in the different indications. In all studies, high levels of PRO-C3 were predictive of poor OS with an increased risk of dying ranging 48%-337%. Conclusions: Type III Collagen pro-peptides (PRO-C3) are prognostic for poor OS across indications. These data suggest that quantifying tumor fibrosis is important for prognostication of patients with cancer and that this can be assessed non-invasively in serum or plasma across indications. [Table: see text]
Evaluation of concordance between blood-based and matched tissue molecular testing in a regional cancer center.
Authors: Darabi Sourat , Zuazo Carlos E , Brodie Taylor , Braxton David R. , Eisenberg Burton Larry , Demeure Michael J.
Published: June 2024

e15062

Background: Next-generation sequencing (NGS) to identify cancer-associated mutations together with an increasing array of targeted drugs with FDA approval allows for matching patients with more effective therapies. Tumor tissue molecular profiling is most often used to identify actionable biomarkers. Blood-based tests to sequence cell-free circulating tumor DNA (ctDNA) are a less invasive approach. Both methods have high specificity and sensitivity and provide insights into actionable biomarkers; however, little is known about how these NGS tests compare when used for the same patient. This study assessed the concordance of paired ctDNA and solid tumor profiling in patients treated in our regional cancer center. Methods: We retrospectively reviewed ctDNA and matching tumor sequencing data test results from CLIA/CAP laboratories between June 2018, and October 2023. The average testing turnaround time was 12.9 days for solid tumor sequencing and 7.7 days for ctDNA testing. We analyzed genomic variants, gene fusions, and microsatellite status in these patient results. The NGS tests for ctDNA included 83 relevant genes which reported single nucleotide variants, small insertions and deletions, splice variants, and limited copy number alterations and gene fusions. The NGS tests for tumor testing were performed on Formalin-Fixed Paraffin Embedded tissue samples using either a 592 relevant gene panel or whole exome sequencing, and either an RNA-based targeted capture panel of 52 genes or RNA whole transcriptome sequencing. Concordance is defined as both ctDNA and tumor testing reporting the same alterations, whereas discordant reports are defined as each reporting different alterations that didn’t overlap between ctDNA and tumor sequencing. Results: A total of 186 patients had both ctDNA and solid tumor sequencing tests done. Cancer types included pancreatic cancer (n = 42), breast cancer (n = 29), lung cancer (n = 28), colorectal cancer (n = 14), and others (n = 73). We observed concordance between tests in 76 (41%) patients in identifying the same genomic alterations. In 103 (55%) patients, we found discordant genomic findings (in the remaining 7 (4%) patients, there were no alterations detected (ND)). Among 103 patients for whom we found discordant findings between the tests, 41 (40%) showed genomic alterations only from the ctDNA test, 46 (45%) had results only from solid tumor profiling, and 16 (15%) had abnormal gene findings from both solid tumor and ctDNA testing but the results did not overlap. Conclusions: Our retrospective analysis comparing sequencing of ctDNA and tumor tissue demonstrated about 40% concordance in genomic findings. Our study indicates that these tests may be complementary. If one accepts a degree of discordance, then testing of ctDNA may be used to identify actionable genomic alterations in the setting of inaccessible or inadequate tumor samples for sequencing.
Dual targeting of αv and α5β1 integrins in triple-negative breast cancer (TNBC).
Authors: Song Fei , Do Kevin , Fein Daniel E.C. , Jahanseir Narges , Joshi Raghav , Mathew Paul
Published: June 2024

e15073

Background: Integrin αv has been implicated in a stem-phenotype of breast cancer (1) and is cross-regulated by fibronectin-binding integrin α5β1 in tumor cells (2). We hypothesized that these integrins are expressed in TNBC cells and dual targeting of αv and α5β1 integrins by a bispecific antibody (BsAbαv/α5β1) will be superior to monospecific integrin antibodies (MsAbs) alone or in combination. Methods: A representative panel of basal-subtype TNBC lines was assembled: MDA-MB-231 (P53 mutated, KRAS mutated), MDA-MB-468 (P53 mutated, PTEN deleted, EGFR amplified) and BT-20 (P53 and PIK3CA mutated, EGFR amplified). Integrin expression was determined by flow cytometry and Western blot in cell lines and by IHC in the residual cancer burden (RCB) of TNBC specimens following neoadjuvant chemotherapy (positive: >2+ membranous, >10% cells). The impact of MsAbs targeting αv and α5β1 integrins alone or in combination versus the BsAbαv/α5β1 was assessed in adhesion, migration, and clonogenic survival. The impact of integrin blockade on nuclear localization of basal markers including FAK, YAP, EGFR and beta-catenin, was assessed. Results: Robust integrin αv expression was present in all TNBC lines with variable α5β1 expression. Marked reduction in α5 and in αv expression following BsAbαv/α5β1 was uniquely demonstrated by Western blot and/or flow cytometry compared to MsAbs, likely due to endosomal trafficking and lysosomal degradation of target integrins as reported previously (Joshi, MCR. 2020). Combined α5 and αv neutralization with MsAbs was superior to individual single agents in blocking adhesion and migration across all TNBC lines studied. However, the BsAbα5β1/αv was superior to combinatorial MsAbs in MDA-MB-468 and BT-20 lines in chemotaxis assays. A significant (>50%) reduction in clonogenic cell survival was noted with the BsAbα5β1/αv compared to MsAbs. BsAbαv/α5β1 blocked nuclear localization of FAK, EGFR, activated beta-catenin, and YAP in MDA-MB-231 uniquely compared to MsAbs alone or in combination. RCB tumor cells were strongly positive for membranous αv [7/9, (78%)]. Diffuse expression of integrin α5 (67%) in alpha smooth-muscle actin-expressing fibroblasts with dual staining for integrin αv was found. Marked CD68+ macrophage and CD3+ infiltration was seen in tumor and stroma of all RCB tumors. Conclusions: Combinatorial α5 and αv integrin targeting with BsAbα5/αvβ1 defines a novel therapeutic strategy with a distinct mechanism of action for further investigation in TNBC. Enrichment of integrins αv and α5 in the inflamed RCB of TNBC tumors and stroma respectively point to a potential role of these integrins in epithelial-stromal interactions and therapeutic resistance. 1. Desgrosellier, Dev Cell. 2014. 2. Joshi, MCR. 2020.
CancerHallmarks.com: A tool for the enrichment analysis of the cancer hallmarks.
Authors: Gyorffy Balazs , Menyhart Otilia , Kothalawala William , Santarpia Libero
Published: June 2024

e15079

Background: The "Hallmarks of Cancer" concept is a useful framework for understanding the fundamental principles that are shared among different types of cancers. However, the lack of agreement on a specific set of genes for these hallmarks makes it difficult to compare and integrate data, leading to varying interpretations of the biological processes involved in cancer across different studies. In this study, we created a consensus set of cancer hallmark genes by combining data from multiple mapping resources and the scientific literature. Methods: We identified 6,763 candidate cancer hallmark genes associated with ten cancer hallmarks by consolidating information from seven studies. There was a considerable variation in the number of genes within the available resources, ranging between 91 and 7,779. We observed a sizable variation in the number of genes connected to each hallmark. The number of genes across cancer hallmarks ranges between 574 („enabling replicative immortality”) and 3,574 ("sustaining proliferative signaling"). The least number of genes are associated with the more recent hallmarks, published in 2011, and the hallmarks „enabling replicative immortality” and „sustaining angiogenesis”. We identified six genes linked to each of the ten cancer hallmarks: TP53, AKT1, MAP2K1, CTNNB1, HRAS, and PIK3CB. Results: As a practical utility of the hallmark concept, we analyzed the enrichment of cancer hallmarks by evaluating genes associated with overall survival. Only genes with a false discovery rate below 0.01, a hazard rate over 1, and a cutoff value over 100 were analyzed to ensure sufficient robustness of the gene sets. The hallmark "genome instability" was most prominent in liver (p=1.0E-05), lung (p=1.0E-05), and pancreatic cancers (p=0.00164). Renal cancer had higher “invasion and metastasis” (p=0.002). Both pancreatic cancer and lung adenocarcinomas were enriched for “resisting cell death” (p=0.00096 and p=0.037, respectively). Overall, while eight cancer types had no significant hallmark enrichment among druggable genes, pancreatic cancers had the most enriched hallmarks (5 out of 10), emphasizing the disease's complexity. These results highlight the usefulness of the hallmark concept as a valuable tool for organizing information, especially in establishing a direct link between genes and biological functions. Conclusions: We have created a unified list of genes linked to cancer hallmarks. We integrated the data into an easily accessible online tool (accessible at www.cancerhallmarks.com) that enables users to identify the most pertinent cancer-associated “candidate hallmark genes” in their own datasets.
A double-blind, randomized, two-arm, single-dose, parallel-group study in healthy participants to compare the pharmacokinetics, pharmacodynamics, and immunogenicity of FKS518 proposed biosimilar to denosumab with the originator (LUMIADE-1 study).
Authors: Sadek Jouliana , Dryja Anna , Szeles Peter , Buccarello Anna Lucia , Monnet Joelle
Published: June 2024

e15097

Background: Denosumab is a RANKL inhibitor used in patients with bone metastases from solid tumors. FKS518 is a proposed biosimilar to denosumab originator product. Biosimilars offer similar efficacy, quality, safety, and favorable economic impact. Once the analytical and functional similarity of a biosimilar has been shown, a comparative human pharmacokinetics (PK) study is conducted to demonstrate that there are no clinically meaningful differences between the biosimilar and the reference product. The present study assessed the PK equivalence of FKS518 to the originator product and provided data on the similarity of the pharmacodynamic, safety and immunogenicity profiles. Methods: A randomized, controlled, double-blind, parallel-group study in healthy volunteers was performed to compare denosumab biosimilar candidate FKS518 versus the originator product. Subjects received a single subcutaneous denosumab 60 mg injection and were followed up for 40 weeks. Primary endpoints PK parameters (Cmax, AUC0-last and AUC0-inf) were natural log transformed and analyzed using analysis of covariance methods; results were then transformed back to the original scale. Safety data were collected throughout the study and analyzed descriptively. Results: 213 male subjects 28-55 years of age, with a baseline BMI between 18.2 and 31.7 kg/m2, were randomized to the originator denosumab (n = 106) or FK518 (n = 107). Results of the primary PK analysis demonstrated bioequivalence between FKS518 and the originator denosumab product since the 90% confidence intervals (CIs) for the geometric least squares mean (LSM) ratios for the 3 primary PK parameters were completely included within the predefined 80.00% to 125.00% equivalence margin (point estimate [90%CI for the ratio]): Cmax: 104.79% [97.04%,113.15%], AUC0-last: 112.29% [104.17%,121.04%] and AUC0-inf : 112.65% [104.27%,121.70%]. Secondary PK parameters were similar as well between FKS518 and the originator: tmax (median: 217.45 hrs. vs. 216.03 hrs. respectively), t1/2 (median: 235 hrs. vs 337 hrs. respectively). The nature, frequency, severity, or resolution of adverse events were comparable across treatments. For immunogenicity, there was no difference between the 2 treatment groups in ADA and Nab incidence at each timepoint. Conclusions: PK equivalence of FKS518 and denosumab originator product was demonstrated with similar PD, safety and immunogenicity profile for the two products. This study adds to the totality of evidence supporting FKS518 as a proposed biosimilar to denosumab.
Efficacy and safety of second-line treatment with surufatinib for anlotinib-resistant radioiodine-refractory differentiated thyroid cancer: An exploratory multicenter study.
Authors: Wang Yang , Qiu Zhongling , Wang Xufu , Han Na , Li Jiao , Zhai Hongyan , Zhou Zhenhu , Gao Rui , Ji Ting , Chen Libo
Published: June 2024

e15127

Background: Radioiodine-refractory differentiated thyroid cancer (RR-DTC) resistant to first-line targeted therapy typically exhibits rapidly progressive nature, lacking standard treatment options. This study aimed to assess the efficacy and safety of surufatinib, targeting vascular endothelial growth factor receptor 1/2/3, fibroblast growth factor receptor 1, and colony-stimulating factor-1 receptor, in RR-DTC patients who had failed anlotinib treatment. Methods: This multicenter study enrolled patients aged ≥ 18 years with at least one measurable RR-DTC lesion (RECIST 1.1), who had previously received anlotinib treatment and experienced disease progression. Primary endpoint was progression-free survival (PFS); secondary endpoints were disease control rate and safety. Results: As of December 25, 2023, 9 patients were enrolled. Median age, 56 (range, 29-69) years; Male, 88.9%; ECOG PS 0, 88.9%; papillary carcinoma/follicular carcinoma, 77.8%/22.2%. Lung (77.8%) and bone (33.3%) were the most frequently involved organs. The duration of anlotinib treatment ≤ 12 months was found in 44.4% patients. All the 9 patients underwent at least one efficacy evaluation with a median PFS of 6.3 months and disease control rate of 88.9%. Subsequent analysis revealed that median PFS extended to 9.20 months in patients younger than 56 years, compared to 5.95 months in older individuals. Patients with over 12 months of prior anlotinib treatment exhibited mPFS of 9.20 months, versus 6.25 months in those with shorter prior treatment durations. Common treatment-emergent adverse events observed were hypertension (44.4%), proteinuria (22.2%), decreased platelets (22.2%), fatigue (11.1%), and hemoptysis (11.1%). Grade 3 adverse events included proteinuria (11.1%), fatigue (11.1%), and hemoptysis (11.1%). Conclusions: In patients with anlotinib-resistant RR-DTC, second-line surufatinib treatment yielded promising efficacy and tolerable toxicity, warranting randomized controlled studies on RR-DTC refractory to first-line targeted therapy. Clinical trial information: ChiCTR2100052020 .
Single-cell and bulk sequencing analysis to evaluate the oncogenic role of PDE3A in pan-cancer and validation in breast cancer.
Authors: Pu Shengyu , Chen Bohong , Zhang Huimin , He Jianjun , Hao Na
Published: June 2024

e15128

Background: PDE3A is involved in critical physiological processes through the hydrolysis of cycle AMP (cAMP) and GMP (cGMP). Its role in the occurrence and progression of cancer has recently attracted attention, but carcinogenic mechanism in pan-cancer has not been fully elucidated. Methods: We explored the expression, prognostic and diagnostic value, mutation, methylation, immune infiltration, and signaling pathway enrichment analysis of PDE3A in pan-cancer by combining multiple bioinformatics databases and single-cell sequencing analysis. Furthermore, we verified the protein and mRNA expression levels of PDE3A and investigated its oncogenic role in the progression of breast cancer (BRCA). Results: PDE3A was expressed at lower levels than normal tissues in most cancers and showed a wonderful prognostic value. PDE3A was strongly correlated with immune cells, especially cancer-associated fibroblast (CAF), and was involved in various vital signaling pathways. Furthermore, PDE3A could facilitate BRCA progression by promoting cells invasion and migration, and it may be associated with the efficacy of endocrine therapy in ER+ BRCA. Conclusions: We provided new insights into the carcinogenic effect of PDE3A in pan-cancer. PDE3A may be a potential biomarker for cancer diagnosis and prognosis, and it is also a promising target for cancer immunotherapy.
The Gundersen Precision Oncology Cohort: A biospecimen resource for mutation characterization and target discovery.
Authors: Kenny Paraic , Feiszt Peter
Published: June 2024

e15184

Background: The generation and widespread sharing of large somatic mutation datasets has revolutionized our understanding of genetic mechanisms driving cancer initiation and progression. For well-understood mutations, this has created the opportunity for individualized therapeutic intervention in a wide range of malignancies. Beyond these well-characterized mutations, there exists a wide range of alterations that remain poorly understood and which require some kind of functional genomics analysis to determine whether such patients are eligible for the targeted therapy used for more typical mutations in that gene, whether these alterations are passenger mutations that don’t affect throughput on the associated signaling pathway, or reflect another more nuanced phenomenon. Whether using ectopic overexpression in cell lines or generation of genetically modified mice, addressing this problem at scale using functional genomics approaches is challenging. A key limitation is the absence of the original tissue context in which that variant co-evolved with other genetic alterations as well as data on the clinical response of the patient to successive lines of therapy. This can be addressed, in part, by the banking of tissue from patient cohorts who have had comprehensive genomic profiling as part of their routine clinical care. For this purpose, we have established the Gundersen Precision Oncology Cohort. Methods: The Gundersen Precision Oncology Cohort is a retrospective collection of tissue specimens, NGS reports and associated clinical data from deceased patients who received cancer care at the Gundersen Health System. The cohort is updated twice per year and banked tissue specimens are available for collaborative research studies. Results: At November 20, 2023 there were data from 780 patients in the cohort, with specimens tested between 2015 and 2023. 54 tumor types were represented, of which lung (n = 196), colorectal (n = 79), pancreas (n = 65), esophageal (n = 58) and breast (n = 46) were most common. The most common diagnostic companies were STRATA Oncology (n = 463) and Tempus (n = 248). Most patients had testing at a single timepoint, but longitudinal testing is available for 13 patients. We present data from some of the studied subgroups, including atypical point mutations in BRAF as well as fusions affecting BRAF and ROS1. Conclusions: This cohort, available for collaborative studies via the Gundersen Cancer Biobank, illustrates the value of banking well-annotated biospecimens to address outstanding questions in oncology.
Enabling accessibility to the CellScape spatial biology workflow for researchers using standard histology protocols and pre-slided specimens.
Authors: Berigan Benton , Braubach Oliver , Ingalls Matthew , Meshik Xenia , Schwarz J. Spencer , Cashion Kayla , Northcutt Adam , Jackson Charles
Published: June 2024

e15185

Background: Solid tumor specimen collection during clinical trials often follows a standard histology protocol, ending in the generation of FFPE sections mounted onto 1 mm histology slides. Though a common sample format for researchers developing tissue-based biomarkers, slides present a data quality challenge for multiplexed spatial biology workflows. Here we present data describing an innovative flow cell design fully compatible with pre-slided tissue specimens. We collected spatial biology data using this slide-compatible flow cell and compared it to the gold standard direct mounting on coverglass. Several rounds of marker staining and imaging demonstrate equivalency in method outcomes including image resolution, signal to noise, and re-interrogation capacity. Methods: FFPE biopsy tissue specimens (tumor punch biopsies and a tonsil section) were mounted on standard histology slides. These specimens were incorporated into flow cells using a new glass coverslip and porting device, CellScape™ Slide, that pairs with a standard histology slide to form a microfluidic chamber. After mounting, each sample was probed for consensus CD markers (CD45, CD3, CD4,CD8,CD20, etc.) through several rounds of iterative immunofluorescence staining using CellScape™ Precise Spatial Multiplexing. The resulting multi-layered whole slide image dataset was then interrogated for spatial resolution and signal-to-noise using digital image analysis. Results: Spatial resolution and signal-to-noise ratio (SNR) were assessed through comparison to data collected from similar samples using the existing CellScape microfluidic chip. The lowest intensity signal collected with at least a 30:1 SNR (acceptable for image analysis) was equivalent or lower in the CellScape Slide preparation (equivalent sensitivity). The highest intensity signal collected was equivalent between the two flow cell designs. Comparing stain quality (specificity, sensitivity) among markers stained through multiple rounds of iterative staining, imaging, and signal remove via photoinactivation showed that the CellScape Slide flow cell is on par with or better than the existing CellScape chip. Further, the usable clear window of the CellScape Slide flow chamber, measuring 16 mm x 44 mm, was found to be greater than twice that of the existing chip. Literature review determined this to also be the largest imaging window available among commercially available spatial biology platforms. Conclusions: Assessment of the CellScape Slide flow cell, innovative in providing access to the CellScape spatial biology workflow for pre-mounted FFPE tissue, determined that the optical performance of the new flow cell is at least on par with the existing flow cell. Further, the usable window is over twice the area, and the new flow cell is fully capable of supporting re-interrogation of the sample.
Preliminary results of atezolizumab and pirfenidone in non-small cell lung cancer.
Authors: Huang Chao Hui , Wick Jo , Komiya Takefumi
Published: June 2024

e15197

Background: Checkpoint inhibitors (CPI) targeting the PD1/PD-L1 axis improved outcomes for stage IV non-small cell lung cancer (NSCLC). Poor response to PD-L1 antibody has been linked to increased cancer-associated fibroblasts (CAF), which interact with cytotoxic T cells (CTLs) similarly to regulatory T cells (Tregs). CAFs, through cytokine production, impair antitumor immunity by affecting CTL function (TGF beta) and inhibiting CTL recruitment into tumors, suggesting that CAFs could be a therapeutic target in CPI-resistant NSCLC. Pirfenidone (P) exhibits anti-fibrotic effects by blocking fibroblast differentiation into CAFs and suppressing TGF beta production and associated signaling pathways/collagens. Atezolizumab (A) targets PD-L1, inhibiting its interaction with PD-1 and B7-1 (CD80) receptors on T cells. We initiated a phase I/II trial to test the combination of AP in recurrent NSCLC after CPI progression. The phase I study aims to determine the maximum tolerated dose (MTD) of AP and assess its safety and tolerability. The phase II study will evaluate its efficacy. The Exploratory Objectives are to measure circulating levels of TGF beta and investigate the expression of CAF-related proteins. Methods: phase I will enroll 3 patients using P at 801 mg PO TID and A at 1200mg IV every 3 weeks. If there is ≤ 1 Dose Limiting Toxicity (DLT), the study will proceed to phase II. If there are ≥2 DLTs, P will be reduced to 534 mg TID. If there is ≤ 1 DLT at this dose, we will proceed to phase II. If there are ≥2 DLTs, then the study will be terminated. Phase II will enroll 16 patients to assess efficacy. The main inclusion criteria are patients with recurrent NSCLC without actionable mutation, after progression with first-line therapy CPI with or without platinum-based doublet chemotherapy, no more than 2 lines of therapy, measurable disease, ECOG 0-2, and adequate organ function. Results: The study enrolled a total of 5 patients (4 males, 1 female), median age of 63.4; 5 were white; 3 patients did not complete the required 21 days of therapy for evaluation (1 non-compliance, 1 disease progression,1 unrelated Urinary Tract Infection); 2 patients were evaluable for toxicity, without DLT; 1 exhibited a partial response to cutaneous metastatic disease, with overall stable disease, 1 had disease progression. Adverse events (AE) in more than 5% of patients included blood disorders, cardiac, gastrointestinal, administration site, metabolic, musculoskeletal, psychiatric, respiratory, and skin disorders. Serious AEs were noted in 2 patients: one experienced gastrointestinal bowel perforation due to disease progression resulting in death, and the other had a respiratory disorder. Conclusions: The combination of AP did not produce unexpected side effects or reach the dose-limiting toxicity threshold. After the phase I portion is completed, the study will proceed to phase II expansion to enroll patients with recurrent NSCLC. Clinical trial information: NCT04467723 .