bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumor sensitization to PARP inhibitors by DNA damaging synthetically lethal acylfulvenes.
Authors: Kulkarni Aditya , Zhou Jianli , Bhatia Kishor , Sharma Panna
Published: June 2023

e15123

Background: 20-30% of solid tumors (ovarian, breast, pancreatic, prostate cancers) harbor some level of deficiency in DNA Damage Repair (DDR) pathways. Such cancers become dependent on alternative DDR pathways to maintain genome integrity, making them selectively vulnerable to DNA damaging agents. The concept of synthetic lethality has been leveraged to treat subsets of such tumors. Clinically, PARP inhibitors (PARPi) have been successful in Homologous Recombination Deficiency (HRD) cancers. LP-100 and LP-184 are acylfulvene (AF) class small molecule prodrugs that are metabolized to active compounds by prostaglandin reductase 1 (PTGR1) that is often elevated in solid tumors. AF-induced DNA lesions are likely selectively processed by repair pathways including BRCA1/2 mediated Homologous Recombination (HR). Methods: Cell Titer Glo based 10 day cell viability assays were used to generate drug IC50 values in vitro. Bliss synergy scores were computed with scores ≥ 10 reflecting synergy. Drug-induced DNA double strand breaks (DSB) were detected and quantified with STRIDE (SensiTive Recognition of Individual DNA Ends) assay in cancer cells. A subcutaneous tumor xenograft model was used to determine tumor volumes in control and drug treated mice. Results: In a BRCA2 mutant ovarian cancer cell line OVCAR3, LP-184 and olaparib co-treatment in vitro showed synergistic effects (Bliss score 10). Similarly in a BRCA2 mutant prostate cancer cell line 22Rv1, LP-100 combination with rucaparib was synergistic (Bliss score 18). 400 nM LP-184 showed a time dependent increase in DNA DSB signals in vitro, 1.7X more so in BRCA2 mutant vs wild type (WT) DLD-1 isogenic colon cancer cell lines up to 24 hours. To validate the hypothesis that AF-induced DSBs remain unresolved if exposed to and further synergize with PARPi in vivo, we tested LP-100 plus olaparib in a DU145 (BRCA2 mutant) prostate cancer xenograft model. LP-100 + olaparib treatment resulted in sustained, durable tumor regression indicating in vivo synergy whereas either agent alone did not produce tumor shrinkage. Day 42 tumor growth inhibition was 66% for 2.5 mg/kg LP-100 alone, 34% for 40 mg/kg olaparib alone and 108% for the combination. Moreover, LP-184 treatment resulted in early onset and complete tumor regression in 7 PDX models of PARPi resistant triple negative breast cancers (TNBC) harboring BRCA1/2 loss whereas olaparib/ niraparib remained entirely ineffective in these. Conclusions: AF response is influenced by tumor DNA repair competence. LP-100 and LP-184 are synthetically lethal resulting from their abilities to cause unresolvable DNA damage where tumor cells express high PTGR1 and are deficient in the HR pathway. LP-184 single agent was superior to PARPi in TNBC and LP-100 combination made an olaparib resistant tumor highly sensitive, thereby facilitating a path to extend the opportunities for both AFs and PARPis and enhance the clinical utilization of PARPi.
The effect of adding a selective FAK inhibitor AMP945 to FOLFIRINOX in a model of pancreatic cancer.
Authors: Burns Christopher , Murphy Kendelle , Cock Terrie-Anne , Devlin Mark , Herrmann David , Timpson Paul
Published: June 2023

e15128

Background: Pancreatic Cancer (PC) is a one of the leading causes of cancer-related death in the world, with a five-year survival rate of ~11%. The majority of patients present with advanced metastatic disease and are treated with chemotherapy. FOLFIRINOX is the standard-of-care treatment for patients in many countries and has been shown to result in a median overall survival of 11.1 months. Focal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase which transduces signaling from integrin receptors, activated through interactions with the extracellular matrix, to regulate cell survival, proliferation and migration, and is often highly expressed in PC. Inhibition of FAK in preclinical models of PC has been shown to sensitize cells to radiotherapy, immunotherapy and chemotherapy. AMP945 is a potent and selective FAK inhibitor currently undergoing a Phase Ib/IIa trial in PC in combination with gemcitabine and nab-paclitaxel (NCT05355298). We now demonstrate that AMP945, in combination with FOLFIRINOX, can significantly improve survival in mouse models of PC, providing strong rationale for clinical testing of this novel treatment combination. Methods: Patient Derived Xenograft (PDX) models of PC in NOGIL2 mice using TKCC10lo patient derived cells were conducted in both subcutaneous and orthotopic formats. Once the tumor was palpable, AMP945 (10 mg/kg, b.i.d.) was administered for four days. On day 8 oxaliplatin (5 mg/kg) and irinotecan (25 mg/kg) were administered, with leucovorin calcium (100 mg/kg) and fluorouracil (25 mg/kg) being dosed on day 9. The mice were monitored until day 12 and the treatment regimen was repeated for up to 20 cycles, or until the experimental endpoint was reached. At the experimental endpoint the mice were sacrificed, the tumor harvested and analyzed. Results: In the subcutaneous model the combination of AMP945 and FOLFIRINOX increased survival by ~35% compared to FOLFIRINOX alone. In the orthotopic model, median survival was extended by ~30% when using the AMP945/FOLFIRINOX combination, compared to FOLFIRINOX alone. In both models there was a significant reduction in tumor size and clear evidence of on-target FAK inhibition in tumors (reduction in pY397FAK levels) in the combination therapy groups. Notably, a significant increase in cleaved caspase-3, a marker of apoptosis, was also apparent in the combination therapy groups. Conclusions: The FAK inhibitor AMP945, given in a pulsed-dosing regimen in combination with FOLFIRINOX, significantly enhances survival in a mouse model of pancreatic cancer by increasing apoptosis and reducing tumor burden, highlighting the potential of this combination in clinical treatment of PC. A clinical study of the combination of AMP945 with FOLFIRINOX is now in planning.
Penpulimab combined with anlotinib therapy in patients with advanced refractory solid tumors: A single-center, observational, prospective study.
Authors: Zhou Shansi , Wang Tao
Published: June 2023

e15129

Background: Antiangiogenesis therapy combined with PD-1/PD-L1 inhibitors has shown excellent efficacy in advanced refractory solid tumors. In advanced NSCLC patients,this conclusion has been confirmed in a number of clinical studies such as WJOG@Be Study and JVDF. A clinical study, KEYNOTE-524/Study116, demonstrated survival benefits from this combination model in liver cancer, and a similar trend was seen in patients in the bile duct cancer study presented.Penpulimab + anlotinib is an important combination immunotherapy. Here we described the results of safety, and clinical efficacy of penpulimab combined with anlotinib in patients with advanced solid tumors, the serum cytokine levels, and peripheral blood T lymphocyte populations were detected simultaneously. Methods: 17 cases with advanced cancers including lung, gallbladder, cholangiocarcinoma, and hepatocellular Carcinoma were treated since January 2022. Patients received a combination of anlotinib (12mg) once daily on day 1 to day 14 (21days as a course) plus anti-PD-1 antibodies every 3 weeks until progression or intolerabletoxicity. Imaging was performed every 6 weeks for thefirst year of therapy. Blood samples were collected from patients prospectively. Serum interleukin (IL-2, IL-4, IL-6, IL-10), tumornecrosis factor-α(TNF-α), interferon-γ(IFN-γ) and circulating immune cell subsets were measured at baseline and after two cycles of treatment via flow cytometry. Results: From January 2022 to December 2022, 17 patients had received the combination therapy of penpulimab plus anlotinib (median age 68.8yrs [range 51-83],52.9% males, 100% ECOG PS 1. Most patients had received frist line treatments for metastatic disease (82.4%). Of 13 pts who have had at least one tumor assessment, the ORR（objective remission rate ）was 35.29% (6 PRs) and DCR was 58.82% (4 SDs). The median PFS was 7.98 months (95% CI: 1.68-NE months).Up to now, eight responders remain in response.The most common adverse reactions included：hypertension was 1/17（5.9%）, Hand foot syndrome was 1/17（5.9%）, loss of appetite was 1/17（5.9%）.There were no grade 3 and above treatment-related adverse events. Conclusions: The combination of penpulimab plus anlotinib as a treatment for advanced refractory solid tumor showed the promising efficacy with a manageable safety profile, thereby suggesting that this combination therapy may be a viable treatment strategy for advanced refractory solid tumor patients.
A novel orally bioavailable small molecular ENPP1 inhibitor (OC-1) for cancer immunotherapy.
Authors: Chen Jiyun , Pujala Brahmam , Gupta Ashu , Ansaria Amantullah , Khana Farha , Lee Elbert , Chakravarty Sarvajit
Published: June 2023

e15136

Background: Ectonucleotide pyrophosphatase/phosphodiesterase I (ENPP1) is a type II transmembrane glycoprotein with nucleotide pyrophosphatase and phosphodiesterase enzymatic activities. Inhibiting ENPP1 activity has emerged as an attractive approach in cancer immunotherapy, based on two distinct mechanisms. One mechanism is the stabilization of cyclic GMP–AMP (2’3’-cGAMP), which binds to stimulator of IFN genes (STING) and functions as an endogenous second messenger to activate the innate immune system; the other is the lowering of adenosine concentrations in the tumor microenvironment. ENPP1 inhibitors have shown promising monotherapy activity and synergy with various targeted therapies in preclinical models. Methods: OC-1 was characterized, for activity and selectivity, in the following in vitro assays: a luminescence-based enzymatic assay; cell proliferation assays; selectivity assays against 15 phosphodiesterases (PDE) based on inhibition of enzymatic activity; and a cytokine release assay in human THP cells and mouse splenocytes looking at changes in IFN-β/CXCL10. Oral bioavailability of OC-1 in mice and rats was studied following oral gavage. Plasma exposures were quantified using standard LC/MS/MS methods. In vivo tumor growth inhibition (TGI) of OC-1 was evaluated in syngeneic mouse tumor models (CT26 and MC38), as monotherapy and in combination with an anti-PD1 antibody. OC-1 was dosed orally in all in-vivo models. Results: We have discovered a novel small molecule inhibitor of ENPP1, OC-1, with a Ki value less than 10 nM in an ENPP1 enzymatic assay. Increase in gene expression of IFN-β and CXCL10/IP10 was observed in OC-1-treated THP1 cells. No inhibition in cell proliferation in various cancer lines was seen up to the highest concentration tested (30 µM). OC-1 was also selective against 15 PDEs with IC50 > 30 µM. The mean oral bioavailability of OC-1 was 72% and 63% in mice and rats, respectively. OC-1 showed single agent activity (20 – 40% TGI) and combination activity of approximately 75% TGI when it was combined with an anti-PD-1 antibody in in vivo efficacy studies with no impact on body weight. Conclusions: In summary, OC-1 is an orally bioavailable ENPP1 inhibitor with promising anti-tumor activity in preclinical models. Its physicochemical properties make it a suitable candidate for clinical investigation.
Clinical and genomic characteristics between DDR germline and somatic mutations in pan-cancer.
Authors: Li Yanchun , Sun Tingting , Zhang Qin , Qi Chuang
Published: June 2023

e15139

Background: The defects DNA-damage repair (DDR) genes would drive tumor formation and are associated with increased genomic instability and tumor mutational burden (TMB) in cancer. Although, alterations of DDR genes are common in tumor, the differences between the germline and somatic alterations are poorly characterized. Methods: The DNA sequencing data of 540 genes from 6174 pan-cancer patients were retrospectively collected and 276 DDR genes were analyzed. The variations were annotated as pathogenic (P), likely pathogenic (LP), and nonpathogenic (NP) according to ACMG (American College of Medical Genetics) guideline. The patients were divided into 3 groups: DDR-germline (P&LP germline variants, N = 766), DDR-somatic group (only P&LP somatic variants, N = 2923) and non-DDR group (NP variants, N = 2485). Results: In pan-cancer, 12.41% patients have germline DDR gene mutations, of which tumors with high germline mutation frequency include ovarian cancer (27/102, 26.47%), glioma (22/136, 16.18%), breast cancer (25/159, 15.72%), genitourinary cancer (17/114, 14.91%), biliary tract cancer (42/284, 14.79%), pancreatic cancer (28/193, 14.51%), colorectal cancer (95/727, 13.07%), head-neck tumors (11/85, 12.94%) and esophagus cancer (16/124, 12.90%). Meanwhile, 47.34% patients have somatic DDR gene mutations, of which tumors with high somatic mutation frequency include esophagus cancer (89/124, 71.77%), colorectal cancer (488/727, 67.13%), endometrial cancer (49/73, 67.12%), head-neck tumors (50/85, 58.82%), pancreatic cancer (111/193, 57.51%), gastric carcinoma (233/415, 56.14%), ovarian cancer (53/102, 51.96%) and biliary tract cancer (138/284, 48.59%). The most commonly germline alterations were found in TP53 (33%), PTEN (14%) and BRCA2 (14%), while in the somatic mutations, TP53 (52%) showed the highest frequency, PTEN (9%) and MSH3 (8%) followed. In pairwise comparisons of the three groups’ actionable mutations, DDR-somatic group was more likely to have alterations in TP53 (OR = 5.68), AKT3 (OR = 5.81) and APC (OR = 1.42) compared with DDR-germline group. Taken non-DDR group as reference, mutations in TP53 (OR = 5.09), PTEN (OR = 7.65), BRCA2 (OR = 5.32) were more common in DDR-germline group, but TP53 (OR = 28.90), PTEN (OR = 7.73), APC (OR = 2.68) in DDR-somatic group. The tumor mutational burden (TMB) was statistically different among the three groups (p < 2.22e−16). The DDR-somatic group exhibits the highest median TMB (5.68) compared with 1.42 in DDR-germline and 2.13 in non-DDR group. Conclusions: The differences in the mutation profile between the DDR-germline and DDR-somatic groups and the distinct actionable genes may indicate the different target-therapy choices. Besides, DDR-somatic group exhibited the highest TMB, which might indicate patients with somatic DDR alterations may better benefit from the immunotherapy.
Clinical characterization of MYC family proto-oncogene amplification in solid tumors from Chinese patients.
Authors: Liu Zhiliang , Han Tiantian , Kong Rongrong , Guo Didi , Wang Mengjuan , Dong Yuwei , Chen Siqi , Fan Jiawen , Deng Wanglong , Ding Ran , Bu Fanfeng
Published: June 2023

e15140

Background: The dysregulation of the MYC family oncogenes ( c-MYC, MYCN and MYCL) play critical roles in tumorigenesis, prognosis and immune escape. MYC inactivation can result in sustained tumour regression and many therapeutic agents that directly target MYC are under development. MYC signaling is associated with tumor cell PD-L1, overall immune cell infiltration. Herein, we explore MYC family proto-oncogene amplification profiles and clinical characterization in chinese solid tumors. Methods: This research comprehensively characterized gene mutations by next-generation sequencing (NGS) in 23990 chinese solid tumors tissues to reveal the prevalence of MYC family proto-oncogene amplification(MYC AMP) and the association with Tumor mutational burden(TMB) and microsatellite instability(MSI). Results: The prevalence of MYC AMP (copy number, CN≥5) in the cohort was 2.1% (504/23,990), in which ovarian cancer (7.6%, 22/289) showed the highest prevalence, followed by breast cancer (6.1%, 26/429), esophagus cancer (5.9%, 17/287). Only one glioma patient(pt) carried co-amplification of MYCN and c-MYC. In 504 MYC AMP pts, c-MYC AMP accounted for 93.7%(472/504), MYCN and MYCL AMP accounted for 6.5%(33/504) in total. The CN was significantly higher in MYCN and MYCL AMP pts than c-MYC AMP pts (22.9 vs 7.6, p < 0.0001). MSI-H showed a lower detection rate in MYC AMP pts other than Non-MYC AMP pts (0% vs 1.2%, p < 0.05). The proportion of TMB-L in MYC AMP pts was similar to Non-MYC AMP pts (90.9% vs. 90.1%, p > 0.05). Conclusions: In totally, 2.1% of chinese solid tumor pts had MYC high level AMP, mainly c-MYC Amp. The CN was higer in MYCN/ MYCL AMP pts than c-MYC AMP pts. In addition, MYC AMP pts tended to have MSS and TMB-L, suggesting that MYC may be a novel target for tumor immunotherapy. MYC inhibitor combines with immunotherapy may be an important direction for the treatment of MYC AMP pts.
Impact of overfixation on actionable biomarkers and checkpoint inhibitor (CKI) targets with immunohistochemistry (IHC) in a patient population with non-small cell lung cancer (NSCLC).
Authors: Gérus-Durand Marie , Gaspo Rania , Tsao Ming Sound , Jean Alexandra , Sallette Jérôme , Burrer Renaud , Finan Amanda
Published: June 2023

e15144

Background: The treatment of patients with advanced NSCLC has become reliant on tissue specimens and biomarkers to help guide appropriate treatment options. Currently, there are numerous lung cancer biomarker-defined patient subgroups, with evidence showing that fixation conditions may alter their expression on FFPE specimens, but little evidence on the ensemble of targetable biomarkers and CKI targets. Methods: We assessed 30 adult patients with primary NSCLC tumors. 25 specimens were properly fixed with neutral 10% formalin using gentle agitation for 24-48 hours, whereas 5 specimens were over-fixed (52-108 hours). We examined expression of actionable and exploratory biomarkers ALK, ROS1, BRAF, EGFR, c-Met, panTRK, HER2 and MEK1. We also examined CKI targets such as PDL1, CD3/CD8, CD3/CD8/FoxP3, PD1, CD3/CD8/PD1. We performed IHC on all slides which were scored by a thoracic pathologist per standard clinical practice. Descriptive statistics were applied to further explore the impact of overfixation on IHC results. Results: Results are shown in the table below. Patients had an average age of 64, were mostly male (22/30) with adenocarcinoma histology (21/30). Specimens were collected in compliance with local rules and regulations and tumor staging was from IB-IV. For the 5 overfixed specimens, multiplex PD-L1 was less expressed when compared to the normally fixed specimens. We also surprisingly observed that cMet and HER2 had stronger expression with overfixed tissue. All other targets had no differences observed in this small patient population. Conclusions: Overfixation of NSCLC FFPE had little to no detrimental impact on most targetable biomarkers, except multiplex PD-L1. We also observed that cMet and HER2 had a slightly stronger expression level with overfixed tissue. As we do not have a reference of the overfixed samples fixed from 24-48 hrs, we cannot be assured that the differences in expression levels are not inherent to the sample itself and independent of fixation time. Additional tests of the same sample fixed over different periods will help to identify the true cause. We propose a consistent approach on tissue fixation time and process as the ensemble of preanalytical conditions is critical in ensuring proper biomarker characterization and validation with IHC. [Table: see text]
Use of CLIA accredited multi-omic platforms to correlate clinical significance of WES/WTS testing with functional protein/phosphoprotein drug target activity.
Authors: Elsey Rachel , Meissner Tobias , Mueller Claudius , Corgiat Brian , Davis Justin B , Hattum Crystal , Spanos William Charles , Starks David , Solomon Benjamin Maurice , Petricoin Emanuel , Lee John H. , Williams Casey B.
Published: June 2023

e15150

Background: Targeted cancer therapies almost exclusively disrupt protein function and phosphorylation, but genomics-based biomarkers underpin the vast majority of oncology molecular profiling. The reverse phase protein array (RPPA) technology has been used in the clinical setting to identify potential drug targets and to monitor the effects of treatment on protein expression and phosphorylation. Here, we investigated the Theralink RPPA assay, a highly sensitive CAP/CLIA accredited protein/phosphoprotein profiling technology, across a pan-tumor cohort of 188 samples which had undergone whole exome sequencing and transcriptomics. Methods: Using the Theralink RPPA assay, 32 key protein and phosphoprotein cancer therapy targets were quantified from formalin-fixed, laser capture microdissected (LCM) tumor epithelium across a pan-tumor cohort of 188 cancer cases. All samples had concomitant CLIA-based whole exome and whole transcriptome analysis performed. Most of these tumors were head and neck cancer (34%), uterine cancer (23%), and ovarian/fallopian tube cancer (17%), with the remaining 26% made up of various cancer types, including skin, breast, lung, brain, and other cancers. Results: Of the pan-tumor cohort, 95% of cases had at least one actionable protein target identified, compared to 86% of cases with clinically relevant genomic alterations. Altered genes and Theralink RPPA assay protein targets were broadly classified into 10 biological pathways. The pathway most frequently identified as actionable via Theralink’s RPPA assay was HER-family protein signaling (85% of cases) with concurrent genomic alterations in HER-family genes found for 9% of cases. In contrast, mTOR signaling was found to be the most frequent clinically relevant genomically altered pathway (48% of cases). When using a clinical breast cancer reference for HER2 protein/phosphorylation levels that is guiding HER2-targeted therapy in the unamplified setting, actionable total and phospho-HER2 levels were found in 82% of the pan-tumor cases, overlapping with ERBB2 genomic alterations in 4% of cases. The gene found to be altered the most was TP53 (101 cases; 54%), which was not part of the targeted Theralink RPPA assay. Conclusions: Our data suggest that quantification of key cancer proteins and phosphoproteins using RPPA extend the identification of actionable drug targets beyond what was found with WES/WTS testing alone. Especially the quantification of total and phospho-HER2 identified a significant pan-tumor subpopulation that may benefit from HER2-targeted therapies, including trastuzumab deruxtecan (T-Dxd).
Multi-omics study of the genomic features and clinical characteristics of hepatic sarcomatoid carcinoma located at different sites.
Authors: Li Yanchun , Guo Wei , Mo Mengjun , Li Yangcan , He Qingqing , Lian Baofeng , Shi Xiaoliang , Lin Guomin , Pang Fei
Published: June 2023

e15153

Background: Hepatic sarcomatoid carcinoma (HSC) is a rare and highly malignant tumor, which accounts for less than 2% of hepatic carcinoma. HSC can occur in the liver (Sarcomatoid hepatocellular carcinoma, SHCC) or in the intrahepatic bile duct (Sarcomatoid intrahepatic cholangiocarcinoma, SiCCA). We aimed to compare the clinical characteristics and genomic features between SHCC and SiCCA. Methods: In this study, we recruited 20 patients who were pathologically diagnosed with HSC from January 2018 to February 2021 in Hunan Provincial People’s Hospital. Whole exome sequencing and PD-L1 expression detected by immunohistochemistry were performed on the qualified tissue samples obtained from18 (14 with SHCC and 4 with SiCCA) of the enrolled patients. Tests were performed in OrigiMed, a College of American Pathologists (CAP) accredited and Clinical Laboratory Improvement Amendments (CLIA) certified laboratory. Fisher test was used to determine the association between two categorical variables, and P< 0.05 was considered statistically significant. Results: A total of 15 cases of SHCC and 5 cases of SiCCA were enrolled in this study, with median age of 59 years old (ranged from 34 to 75 years old). Among them, 15 (75%) were diagnosed as AJCC stage III-IV. Compared with SiCCA, we observed a significant higher proportion of undifferentiated samples in SHCC ( P< 0.001) (Table 1). The most commonly mutated genes of HSC included TP53 (64% in SHCC and 75% in SiCCA) and TTN (43% in SHCC and 100% in SiCCA). The most commonly mutated genes in SHCC also included TERT (43%), CCDC168 (36%), and LATS1 (36%), and that in SiCCA also included CDKN2A (50%), KRAS (50%), and CDKN2B (50%). All these genes were mainly enriched in cell cycle and ERBB pathway. In addition, amplifications of HYDIN, C4A, LRRC37A and deletions of CDKN2A, NTRK2, NTRK3, NRG1, PAX3, OR4K5, TPP2 and KRTAP4-6 were detected in the SHCC cohort, and amplifications of ZDHHC11, PMS2 and deletions of NRG1 and CDKN2A were detected in the SiCCA cohort. We also assessed TMB and PD-L1 status among these two cohorts. We found that 86% (12/14) of SHCC and 100% (4/4) of SiCCA had TMB of lower than 10 mutations/Mb. The expression of PD-L1 was positive (CPS > = 1) in 86% (12/14) of SHCC and 75% (3/4) of SiCCA. Conclusions: We are the first to discover the significant differences in tumor differentiation between SHCC and SiCCA. We also profiled the mutational characteristics, TMB status, and PD-L1 status of HSC, shedding light on the precision medicine of HSC. [Table: see text]
Pattern of CD8+ tissue resident memory T cells between local recurrence and distant metastasis in nasopharyngeal carcinoma.
Authors: Kam Ngar Woon , Lau Cho Yiu , Lai Syrus Pak Hei , Che Chi Ming , Lee Victor Ho-Fun
Published: June 2023

e15161

Background: CD8+ T lymphocytes are believed to be the major anti-tumor immune subsets. Consequently, most cancer immunotherapeutic approaches seek to amplify cytotoxic T lymphocytes specific to malignant cells. A recently identified subpopulation of memory CD8+ T cells, named tissue-resident memory T (Trm) cells, persists in non-lymphoid tissues, including solid tumors. This T-cell subset is considered an independent memory T-cell lineage with cytotoxic features. However, their function, spatial location and influence on disease-related outcomes (local recurrence [LR], distant metastasis [DM] in nasopharyngeal carcinoma (NPC), a disease entity characterized by substantial immune infiltrate, remain poorly understood. We aim to close this gap through investigation on the abundance and spatial localization of CD8+ Trm in relation to tumor site (primary or recurrence [LR and DM]). Methods: This retrospective cohort analysis based on 20 NPC patient samples: primary (initial diagnosis; [I; n = 19], recurrence ([LR; n = 14] and [DM; n = 10]). We accessed the spatial localization in tumor tissues within tissue-segmented classifications of either cancer islands (pan-cytokeratin–positive [CK+] areas) or stromal areas (CK– areas). Tumor infiltrating lymphocytes (TILs) among patients from I, LR and DM group was quantified through multiplexed 5-colour opal staining using biomarkers against CD8, CD103, IL17, CK plus DAPI (nuclear cell counterstain). Results: While total CD8+ TILs (total tumor) as well as their spatially localized cell densities (tumor islet [TI] vs. tumor stroma [TS]) did not show any significant changes among I, LR and DM patient groups, a higher proportion of CD8+ TILs in the TI co-express the CD103 molecule, an indication of CD8+ Trm (CD103+CD8+) infiltration (mean CD103+CD8+/CD8+ in TI: 32.1% in I group, 44.8% in LR group, 46.0% in DM group; mean CD103+CD8+/CD8+ in TS: 19.0% in I group, 26.2% in LR group 26.2% in DM group). Notably, we observed that such augmentation only reached statistically significant within TI from patient in LR group (p = 0.0451). Furthermore, we found that CD8+ Trm in the LR group expressed higher level of cytokine IL-17, which was ~2.1-fold (p = 0.0788) and 3.6-fold upregulated (p = 0.0285) as compared to I and DM, respectively. Conclusions: Cancer islet–localized CD8+ TILs are composed of CD103+ Trms, which make up nearly 45% the total CD8+ T cell population within LR tumors of NPC patients, highlighting spatial localization of CD103+CD8+ Trms as a key phenotype of CD8+ TIL subset. The CD8+ Trm expressed with increased IL17 suggests that enrichment of this T-cell subset might exerts cytotoxic functions. Our results demonstrate that CD8+ Trm had a greater likelihood as an indicator of LR risk for NPC patients and help identify patient cohorts likely to benefit from immunotherapy.
Comprehensive multigene profiling impact on clinical decisions in patients with advanced cancers: A multicenter, retrospective analysis.
Authors: Kuznetsova Olesya , Ivanov Maxim , Tryakin Alexey , Lebedeva Alexandra , Ignatova Ekaterina , Fedyanin Mikhail
Published: June 2023

e15163

Background: Comprehensive molecular profiling (CMP) via next generation sequencing (NGS) is currently applied in clinical practice and has enabled the detection of biomarkers of response to targeted treatment. Given the lack of the data about optimal range of analyzed alterations, the clinical usefulness of CMP has not been established. To address the challenges in incorporating genomics in treatment of oncology patients, we conducted a retrospective study. Methods: Preliminary analysis of NGS reports and clinical data of patients with advanced solid tumors was performed. CMP was conducted in 261 cancer patients (non-small cell lung cancer - 30,6%, colorectal cancer - 24,8%, pancreatic cancer - 7,4%, breast cancer- 6,2%; 59,3% were female, median number of prior systemic treatment lines was 2 (0-12)). In the present study, tumor molecular profile analysis was performed using 160 - 400 gene NGS panels, containing the majority of clinically significant genes for cancer treatment selection. We assessed the percentage of patients who were recommended to receive molecularly-matched therapy (MMT) after CMP, immunotherapy (IT) distribution of recommended therapies according to ESCAT tiers, median progression-free survival (PFS) – ratio ( = PFS after CMP / PFS before CMP, PFS2/PFS1), median PFS2. Results: MMT after CMP was recommended for 55 (21,1%) patients. ESCAT tiers could be assessed in 48 MMT patients (87%). For 31% of patients, administered therapy was considered as ESCAT I tier recommendation (data on mPFS2/1, mPFS2 not available). ESCAT tiers II, III, IV and IT administration were considered in 31, 8, 8 and 22% of patients respectively (detailed data is presented in Table 1). Most commonly altered genes were TP53 (53%), RAS (29%), APC (18%), CDKN2A/B (16%), PIK3CA(14%), NF1 (10%), BRCA1/2 (9%). Positive predictive factors for MMT after CMP were MSI-high status (HR 0,13, 95% CI 0,03 - 0,67), presence of EGFR alteration (HR 0,32, 95% CI 0,12 - 0,86). Presence of RAS mutation was considered a negative predictive factor (HR 2,78, 1,07 - 7,26). Conclusions: Available NGS panels offer a wide range of analyzed alterations that are not feasible in clinical practice. Most of the alterations that can cause impact on treatment plan can be detected with other methods than NGS. Further studies are needed to determine optimal range of alterations depending on tumor site. [Table: see text]
Comprehensive genomic analysis for homologous recombination deficiency in pancreatic cancer.
Authors: Jung Kwangrok , Jung Jae Hyup , Ahn Jinwoo , Kim Bomi , Lee Jong-Chan , Kim Jaihwan , Hwang Jin-Hyeok
Published: June 2023

e15168

Background: Although homologous recombination deficiency (HRD) has been heavily studied in oncology, HRD in pancreatic cancer remains poorly understood beyond germline alterations in BRCA1, BRCA2 and PALB2. Methods: We performed whole exome sequencing (WES) on 205 patients with pancreatic cancer. Germline and somatic mutations in HRD genes were evaluated: (1) canonical HRD mutation ( BRCA1, BRCA2 and PALB2); and (2) non-canonical HRD mutation ( ATM, ATR, ATRX, CHEK2, BAP1, BARD1, BLM, BRIP1, NBN, RAD51 genes, RAD54 genes, FANC genes and ERCC genes). In addition, signature-based HRD classifiers were also applied: (1) the genomic instability score (GIS); (2) substitution base signature 3 (SBS3); and (3) small insertion and deletions 83B (ID83B). The frequency of HRD mutations and signatures-based HRD in pancreatic cancer were evaluated and their clinical utility were assessed by confirming the association with the response to platinum-containing chemotherapy. Results: Among the 205 patients, 41 patients (20.0%) were identified to harbor HRD mutations. Germline and somatic HRD mutations were identified in 19 (9.3%) and 24 (11.7%) patients, respectively. Mutations in canonical genes and non-canonical genes were found in 12 (5.9%) and 37 (18.1%) patients, respectively. Using signature-based HRD classifier, additional 45 (22.0%) patients were determined to be HRD. Among patients who received first-line FOLFIRINOX ( N = 113), canonical HRD mutation and signature-based HRD group showed higher response rate than non-canonical HRD and non-HRD group (75.0% and 57.1% vs. 31.3% and 33.3%, respectively). In the survival analysis of patients treated with FOLFIRINOX, canonical HRD and signature-based HRD group tended to have better overall survival than non-HRD group. Conclusions: Comprehensive genomic analysis could identify a significant number of HRD pancreatic cancer beyond germline BRCA1,2 and PALB2 mutation, and it would be helpful to choose the appropriate chemotherapy regimen.
Carcinoembryonic antigen (CEA) expression in human tumors: A tissue microarray study on 15,413 tumors.
Authors: Jansen Kristina , Kornfeld Lara , Lennartz Maximilian , Blessin Niclas C. , Höflmeyer Doris , Dwertmann Rico Sebastian , Kind Simon , Viehweger Florian , Fraune Christoph , Gorbokon Natalia , Hube-Magg Claudia , Menz Anne , Burandt Eike C. , Sauter Guido , Simon Ronald , Kluth Martina , Steurer Stefan , Marx Andreas H. , Minner Sarah , Bernreuther Christian
Published: June 2023

e15181

Background: Carcinoembryonic antigen (CEA) is a cell surface glycoprotein which is overexpressed in various cancers but reported prevalence data vary considerably for many tumor types. Because of its overexpression in various cancer types and the limited expression in normal tissue CEA has become a potential drug target with several potential drug candidates currently being evaluated. CEA is also shed into the blood stream, so that CEA measurement in the serum is used as a tool for early detection and recurrence monitoring of cancer. Methods: To comprehensively determine CEA expression in normal and neoplastic tissues, a tissue microarray containing 15,413 samples from 120 different tumor types and subtypes as well as 76 different normal tissue types were analyzed by immunohistochemistry. Results: CEA was detectable in 65 (54.2%) of 120 tumor categories including 49 (40.8%) tumor types with at least one strongly positive case. CEA positivity was most common in colorectal adenomas (100%) and carcinomas (98.7%), other gastrointestinal adenocarcinomas (61.1%-80.3%), medullary carcinomas of the thyroid (96.3%), pulmonary adenocarcinoma (73.7%), mucinous carcinomas of the ovary (79.8%) and the breast (43.2%), squamous cell carcinomas of various sites (30.2%-69.1%), and small cell carcinomas of the lung (64.3%), the urinary bladder (38.9%), and the prostate (50.0%) as well as non-invasive papillary urothelial carcinoma pTa G3 (33,6%), pTa G2 high-grade (25,0%) and pTa G2 low-grade (5,7%). High CEA expression was linked to high grade tumors (p < 0.0001) and invasive growth (p < 0.0001) in urinary bladder cancer as well as estrogen receptor positivity (p = 0.0005) and HER2 positivity (p = 0,0158) in invasive breast cancer of no special type. In 1.250 colorectal adenocarcinomas, reduced CEA expression was associated with mismatch repair deficiency (p < 0.0001) but not with pT and pN stage. CEA expression level was unrelated to clinico-pathological tumor parameters in adenocarcinomas of the pancreas and the stomach, endometroid endometrium carcinoma as well as in serous and endometroid carcinomas of the ovaries. Conclusions: In summary, CEA is abundantly expressed in a broad range of epithelial neoplasms. Our data thus identify various tumor entities where CEA positive cancers might best benefit from CEA serum monitoring and anti-CEA therapies.
Antiproliferative activity of marine-derived oligopeptides combined with androgen receptor inhibition in preclinical prostate cancer models.
Authors: Currie Crawford , Bjerknes Christian , Hermansen Erland , Framroze Bomi
Published: June 2023

e15186

Background: Recent cancer statistics reveal a sustained increase in the incidence of prostate cancer (PC) of 3% per year. More than half of this increase is attributable to advanced disease. Advanced PC remains largely incurable, owing to inevitable progression. Escalation of therapy can restore tumor control, but significant sequelae may result. Anticancer peptides of marine origin appear intrinsically target specific and may constitute a novel approach to improve the therapeutic sensitivity of resistant cancers. Recently published in vitro work showed that a marine soluble protein hydrolysate (SPH) potentiated the anticancer effect of androgen-receptor inhibitors (ARis) in PC cells. SPH also induced transcriptional upregulation of ferritin heavy chain 1 (FTH1) in LNCaP and PC3 cell lines. Given the growing recognition that iron dysmetabolism plays a key role in PC, we applied the findings to this current study. Methods: Eight candidate FTH1-modulatory peptides (FT-001 to -008, all 7- or 8-mer peptides) were identified in the SPH peptide mix and subsequently sequenced and synthesized. Antiproliferative effects of individual FT peptides were assessed in combination with ARis (bicalutamide/BIC or enzalutamide/ENZ) in phenotypically distinct PC cell lines (LNCaP, PC3, VCaP, ENZR-VCaP) and compared to the inhibitory effect of SPH-ARi combination or ARi monotherapy. Results: FT-002 or -005 (10 μM concentration), showed significantly greater inhibition of LNCaP and PC3 cell viability when co-administered with BIC IC50 (0.4 μM & 10 μM respectively), than SPH (160 μM) with BIC or BIC monotherapy (p < 0.001). Similarly, either peptide combined with ENZ IC50 (1.4 μM) exhibited significant antiproliferative effects on androgen-sensitive VCaP cells when compared to SPH (160 μM) & ENZ co-treatment (p < 0.01) and ENZ monotreatment (p < 0.001). Both peptides (10 μM) produced a similar pattern of results in ENZ-resistant VCaP cells when treated with ENZ at its corresponding IC50 (47 μM) (p < 0.001). Differential gene expression profiling of all tested cells showed a consistent and significant upregulation of FTH1 and concomitant downregulation of transferrin receptor (TFRC) gene expression of greater than 2-fold for FT/ARi therapy. Conclusions: FT-002 and -005 synergistically combine with ARis to significantly attenuate PC tumor growth in vitro. Gene expression profiling implicates peptide-mediated FTH1 and TFRC modulation in the mode of action to interfere with tumor cell iron metabolism. Novel research perspectives implicate ferritin and FTH1 expression levels in prostate carcinogenesis and tumor progression. Further research in xenograft models is warranted, in order to validate the current findings and further investigate the cellular pathways responsible for the observed anticancer activity seen with FT-002 and FT-005.
Marketing approvals for cell and gene therapy products in oncology: Current review of EMA and FDA approvals.
Authors: Chandaria Rehma , Sripathirathan Kumar , Song Yun Jeong , Learn Chris A.
Published: June 2023

e15189

Background: Cell and gene therapies (CGT) are emerging as potentially game-changing treatments for relapsing and refractory cancer. To date, there are 9 CGTs approved for different cancers across the US and EU, majority focused on autologous CAR-T cell therapies for hematological cancers. In addition, there are ~1500 CGTs in preclinical and clinical development for treatment of cancers utilizing diverse approaches. The risks involved with CGTs differ from small molecule and biologics, and therefore the data required to demonstrate their safety are also different. To get an insight into what regulatory agencies are looking for demonstrating safety for anticancer CGTs, we analyzed the supporting nonclinical studies for the currently approved therapies. Methods: We reviewed of the European Public Assessment Reports (EPAR) and FDA Summary Basis for Approval (SBA) documents for the 9 approved anticancer CGTs. In particular, we looked into approaches that were accepted by the regulatory agencies and the justification provided. Results: Of the 9 approved CGTs included in the review, there were 6 autologous CAR-T cell therapies for the treatment of different hematological malignancies, 1 PBMC-based cell therapy for prostate cancer, 1 gene therapy treatment for bladder cancer, and 1 oncolytic virus for melanoma. Of the different cell therapies, only for Carvykti was a non-human primate study conducted with autologous CAR-T cells. The absence of toxicity studies for the other cell therapies was justified by the lack of relevant animal models since autologous human cells do not work in immune competent animals. Safety data was gathered along with pharmacology studies and clinical studies. As stated in ICH guideline S9, genotoxicity and carcinogenicity studies are not essential to support clinical trials for therapeutics intended to treat patients with advanced cancer. However, there is a theoretical risk of insertional mutagenesis with gene therapies. This was addressed with the approved gene therapies using studies for integration site analysis and IL-2 independent growth, together with review of published literature and clinical data. Conclusions: The risks associated with each cell and gene therapy are unique. As the science continues to evolve, together with new technologies and approaches, existing regulatory guidance may not cover the nonclinical study requirements for all products. It is therefore crucial for drug developers to take a risk-based approach in determining the need for appropriate nonclinical safety studies. The application of 3Rs to reduce animal use is widely encouraged and therefore regulatory agencies accept lack of non-relevant animal models as well as other science-based justifications for not conducting conventional animal studies.
Influence of age, sex and tumor type in the likelihood of inclusion in early clinical trials.
Authors: Vieito Maria , Rojas Katerin Ingrid , Berché Roger , Viaplana Cristina , Lostes Julia , Galvao Vladimir , Alonso Guzman , Oberoi Honey Kumar , Brana Irene , Oliveira Mafalda , González Macarena , Felip Enriqueta , Saura Cristina , Carles Joan , Macarulla Teresa , Garralda Elena , Tabernero Josep , Saavedra Omar
Published: June 2023

e15193

Background: Inclusivity in early drug development is essential for achieving consistent and accurate results. Sex-specific differences in biology and pharmacology should be explored as early as possible during clinical development. Lack of inclusion of elderly patients underestimates the risk of toxicity due to comorbidities and polypharmacy, but adolescents and young adult patients (AYAs) are even less represented in clinical trials, leading to slower improvements in survival. Methods: Patients referred for a first consultation to Vall de Hebron Institute of Oncology Early Drug Development Unit between 2019-2021 were analyzed. Clinical variables included: age, sex, primary tumor location, clinical trial inclusion and type of clinical trial. Results: On a total of 1218 pts, a total of 580 (48.53%) were included in at least 1 early clinical trial. The median age of the overall population was 61 y(ranging from 22 to 92), with no differences in pts included (62y) or not(60y) in clinical trials. Elderly patients ( > 75y) were 9.1% of the population referred and achieved a similar rate of inclusion (46.90% vs 49.44%) while AYAs(18 to 29-year-old) represented only 1.47% of pts referred and were only matched to a clinical trial in 33.33% of cases. (Fisher’s p-value = 0.06). Equal proportion of males and females were referred to clinical trials, with the exception of elderly patients (60.17% male) and AYAs(61.11%). the overall population). In the overall population the most common primary tumor locations were colorrectal (24.38%), other gastrointestinal tumors (18.14%) and breast cancer (10.91%). The most common primary tumor location in patients > 75y was colorectal(28.82%), other gastrointestinal tumors(19.81%) and head and neck(14.41%) vs sarcoma(38.88%), CNS(27.77%) and other gastrointestinal(11.1%) in AYA patients. Inclusion rates depending on the type of tumor ranged from 63% to 23%. Patients with genitourinary and gynecologic cancers had a statistically significant higher likelihood of being included in a clinical trial whereas sarcoma and primary CNS tumors had lower chances of being included in a clinical trial. In a binomial multivariant model the only factor predictive of inclusion in a early clinical trial was tumor type, whereas, sex, age group and availability of molecular profiling, was not statistically significant. Conclusions: Elderly patients with cancer and AYAs were referred less frequently to our unit that the overall prevalence of cancer in those populations and in the case of AYA pts the rate of inclusion lagged behind, driven mainly by the tumor types more common in each population.
Aluminium phthalocyanine–mediated photodynamic therapy promotes antitumour activity in oesophageal cancer stem cells.
Authors: Didamson Onyisi Christiana , Chandran Rahul , Abrahamse Heidi
Published: June 2023

e15194

Background: Photodynamic therapy (PDT), a non-invasive treatment modality, has emerged as an effective treatment measure for various cancers, including oesophageal cancer. PDT has attracted much attention in the field of oncology due to its low toxicity profile and the selective accumulation of the photoactivated agent in the tumour cells. However, the therapeutic effect of Aluminium (III) Phthalocyanine Chloride Tetra sulfonic Acid (AlPcS4Cl) mediated PDT on oesophageal CSCs is limited. In this study, we evaluated the cellular localization of AlPcS4Cl and its PDT anticancer effects on oesophageal CSCs. Methods: An in vitro experimental design was employed in this study. Oesophageal cancer (HKESC-1 cells) was grown, and the CSCs were isolated using the magnetic-activated cell sorting (MACS). The isolated CSCs were characterised using a flow cytometry, Hoechst efflux side population test, and immunofluorescence. The CSCs were maintained in a serum-free culture medium and incubated at 37° C, 5% CO2 and 85% humidity. The cells were grouped into control and experimental groups. The CSCs received two-fold increasing concentrations of AlPcS4Cl and were exposed to irradiation at 673.2nm wavelength using a diode laser. After 24 hours post-PDT, the anticancer activities of AlPcS4Cl on oesophageal CSCs were examined. The MTT cell viability assay was used to determine the 50% inhibitory concentration (IC50), light microscopy for morphological changes, and adenosine triphosphate (ATP) assay for cell proliferation, lactate dehydrogenase (LDH) assay for cellular toxicity. While cellular localization of AlPcS4Cl was examined using fluorescent microscopy. All experimental and control cells were conducted in six biological replicates (n = 6). Results: Results were compiled, and statistical analysis was performed using GraphPad Prism (v5). The mean values of experimental groups were compared with the mean value of the control cells. One-way ANOVA was employed, and a p-value of less than 0.05 was considered statistically significant. Findings from cellular localization showed that the AlPcS4Cl is localized in the mitochondria and lysosomes, indicating a probable cellular death pathway. The dose-response of AlPcS4Cl on oesophageal CSCs showed an IC50 value of 4µM, suggesting the photosensitizer is efficient for the treatment of CSCs at low concentrations. The CSCs exposed to PDT showed remarkable morphological distortion, high cytotoxic activity, decreased viable cells and significant antiproliferative potential. Conclusions: This study showed that AlPcS4Cl promote PDT anticancer effects on oesophageal CSCs and could serve as an efficient therapeutic option for eradicating oesophageal CSCs.