bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrated survival analysis.
Authors: Chiu Vi Kien , Lerolle-Chiu Adele
Published: June 2023

e14695

Background: Clinical trial efficacy may be determined by the primary endpoint(s) of median progression free survival (mPFS) and/or median overall survival (mOS) from the Kaplan-Meier estimate, and secondary endpoints of overall response rate (ORR) and duration of response (DOR). In the treatment of metastatic cancers, single agent immunotherapy tends to have lower ORR and longer DOR in comparison to chemotherapy or targeted tyrosine kinase inhibitor therapy, which tends to have higher ORR and shorter DOR. This leads to intersecting or crossing Kaplan-Meier survival curves, which are harder to interpret. In immunotherapy trials, mPFS is a poor surrogate for mOS, and both may underrepresent the clinical benefit over time. Methods: We aimed to develop novel metrics that more effectively represent the Kaplan-Meier estimate. The graphical curve of the Kaplan-Meier estimate may be represented by the equation as a function of time. The integration of f ( x), which mathematically represents the area under the curve and clinically the cumulative or integrated survival benefit with time, is: ∫0t f( x) dx = F(t) − F(0). Kaplan-Meier curve equation with constants C and A may be approximated by: ∫0t C e-A x dx = C e-A t ]0 t = C( e-A t – 1). The average integrated clinical benefit with time is: 1/ t ∫0t f( x) dx = [ F( t) − F(0)]/ t. Results: The Phase 3 KeyNote-061 trial of metastatic gastric cancer treated with single agent Pembrolizumab immunotherapy versus Paclitaxel chemotherapy was use for integrated PFS and iOS calculation. The Kaplan-Meier curves intersected or crossed in KeyNote-061 trial. We show the established ORR, mPFS, mOS, DOR. Integrated PFS (iPFS) and iOS were calculated with greater increase in iOS and much less decrease in iPFS for Pembrolizumab than Paclitaxel treatment. Conclusions: Integrated survival is a novel analysis that integrate the Kaplan Meier estimates and better represent the entire median survival and drug DOR may be combined into a single useful metric for overall clinical benefit. [Table: see text]
Influence of CD47 on CD8+ T-cell and tumor prognosis based on pan-cancer analysis.
Authors: Liang Hongxin , Yao Lintong , Xie Daipeng , Qiu Hongrui , Lin Zenan , Li Hao , Leng Jinhang , Wang Huili , Jiao Xiuwei , Qiu Liyun , Zhou HaiYu
Published: June 2023

e14699

Background: CD47 was an immune checkpoint, constituting the CD47/SIRPα axis, sending the 'Do not eat me' signal to macrophages, hereby promoting immune escape of tumor cells in the tumor immune microenvironment(TIME). The role of CD47 has been proved affected immunotherapy efficacy and patient outcomes. However, how it affects TIME, specifically on CD8+T-cell in pan-cancer, has not been fully illustrated. Therefore, we aimed to identify it, which could help us to gain insights into potential anti-cancer treatment. Methods: We used a lot of R package and computational tools to analysis of differential gene expression, prognosis, immunoinfiltration, pathway enrichment and Correlation. Single-cell sequencing and multiple immunofluorescence staining were uesd to verify our hypothesis. Data was based on 17,382 samples given by 948 healthy individuals from GTEx, 39 cancer types from TCGA, 79 databases and 2045746 cells from tumor patients and healthy donors in TISCH, and our real-world sample. Datasets were normalized for estimation of the TME components at both the single-cell and annotated cluster levels. Our study investigated the role of CD47 in prognosis and TIME, mainly from the view of adaptive immunity. Results: CD47 was expressed in nearly all cancer types, associated with worse prognosis in various cohorts in pan-cancer(p＜0.05) compared to the normal tissues. Then, we found CD47 was better correlated with tumor immune evasion through T-cell infiltration compared with T-cell rejection in multi-cancers. And we detected high CD47 expression with high CD8+T-cell infiltration had a worse prognosis. So we hypothesis high CD47 could linked with T-cell exhausted. Then, we investigated that genetic alterations and oncogenic characteristics of CD47 were concomitant with the CD8+T-cell exhausted by analysis of pathway and gene in mulit-cancer especially squamous-cell carcinomas. Finally, we proved CD47 synergy with CD8+T-cell exhausted genes in LUSC and ESCA by single-cell analysis and multiple immune fluorescence staining. Conclusions: CD47 is highly associated with CD8+T-cell exhausted and may induce cancer immune evasion leading to a worse prognosis. Thus, CD47 and T-cell exhausted correlated genes may serve as dual antigenic targets and provide insights into potential immunotherapy.
Efficacy and safety of penpulimab-containing regimens for advanced malignant solid tumors that have failed immunotherapy: A real-world study.
Authors: Zhou Juying , Xu Chao , Zhang Lu , Ma Chenying
Published: June 2023

e14700

Background: Immuno-checkpoint inhibitors (ICI) have shown good anti-tumor efficacy and survival benefits in a variety of solid tumors, such lung cancer, gastric cancer, esophageal cancer, head and neck squamous cell carcinoma (HNSCC). ICI have rewritten the guidelines for multiple tumor types. However, there are still a large number of patients who are ineffective or initially effective after use ICI, and subsequently develop ICI resistance. For these patients, how we choose further treatment is currently uncertain clinically. Some studies suggest that even if the treatment of PD - (L) 1 is ineffective or progresses, it can still bring a certain degree of benefit to re-challenge with other target ICI. This study was to evaluate the efficacy and safety of the patients with advanced malignant solid tumors with the regimen of penpulimab. Methods: This is a non-interventive registration study. Eligible participants must be ≥ 18 years old and have proven malignant tumors that are ineffective, fail for immunotherapy and have no other standard treatment. Patients were treated with penpulimab alone or in combination with chemotherapy or anti-angiogenic agents until progression or unacceptable toxicity. The primary endpoint is objective response rate (ORR). Secondary endpoints were disease control rate (DCR), progression free survival (PFS), overall survival (OS) and safety. Results: Form Sep 2021 to Feb 2023, 28 patients (median age 59.5 years, male 71.4%）were recruited, including 9 pts with NSCLC, 5 pts with HNSCC, 4 pts with Nasopharyngeal carcinoma (NPC), 3 pts with esophageal cancer and 7 with other tumor types. Among all, 1 achieved confirmed PR, 21 achieved SD, the ORR was 3.6% (1/28) and the DCR was 78.6% (22/28). The median treatment duration was 9 months. Among the 9 pts with NSCLC, the median treatment duration was 5.6 months and the DCR was 77.8%. Treatment-related adverse events (TRAEs) of grade 3 were observed in 5 (17.9%) of the 28 patients. The most common TRAEs were decreased lymphocyte count (50%, 14/28), anemia (50%, 14/28), hypoalbuminemia (39.3%, 11/28), hyponatremia (25%, 7/28) and hypercholesterolemia (25%, 7/28). Conclusions: Penpulimab-containing regimens for advanced malignant solid tumors that have failed ICI have shown significant efficacy and a favorable safety profile.
Immune-related hypophysitis: An analysis of a heterogeneous entity.
Authors: Nanah Abdel Rahman , Abdeljaleel Fatima , Ogbue Olisaemeka , Unlu Serhan , Sorathia Sharmeen , Daw Hamed
Published: June 2023

e14704

Background: Immune-checkpoint inhibitors (ICIs) have led to revolutionary breakthroughs in the management of many malignancies. With the increasing use of ICIs, immune-related adverse events (irAE) have emerged and led to significant challenges and limitations. Immune-related hypophysitis, a potentially life-threatening irAE, has limited data on its diagnosis, clinical findings, and management. This was our incentive to characterize our single-institution experience with immune-related hypophysitis over the past 5 years. Methods: We retrospectively studied a cohort of adult patients who received ICIs at the Cleveland Clinic Foundation between 2017-2022. Our search was done by utilizing an outpatient oncology pharmacy database. Analyzed ICIs included atezolizumab, nivolumab, pembrolizumab, ipilimumab, avelumab, and durvalumab. The occurrence of immune-related hypophysitis was as determined by an endocrinologist or oncologist. Results: 6,615 patients received ICIs from 2017-2022, of which 27 patients were diagnosed with hypophysitis. Immune-related hypophysitis occurred with ICI monotherapy in 44.4% and combined therapy (ipilimumab and nivolumab) in 55.5%. The most commonly implicated agents were ipilimumab (74%), followed by nivolumab (59.3%), pembrolizumab (18.5%), and durvalumab (3.7%). In patients receiving ipilimumab-nivolumab, hypophysitis was detected after a median duration of 65 days (range: 29-163 days) from ICI initiation, compared to 58 days (38-112) with ipilimumab alone and 155 days (8-294) with pembrolizumab. Hypophysitis was grade 1 in 18.5%, grade 2 in 29.6%, grade 3 in 37%, and grade 4 in 14.8% of all patients. No hypophysitis-related deaths occurred. Signs and symptoms included constitutional symptoms (fever, anorexia, myalgia) in 88.89%, severe/intractable headaches in 40.7%, gastrointestinal complaints in 29.6%, dermatologic findings in 7.4%, visual symptoms in 7.4%, and syncope in 3.7% of all patients. Lab findings included abnormalities of TSH in 77.78%, ACTH in 70.60%, FSH and/or LH in 52.63%, and a positive ACTH stimulation test in 69.3%. Imaging findings were present in 62.5% of patients, and most commonly revealed pituitary gland enlargement, enhancement, or stalk thickening on brain/pituitary MRI. Treatment included glucocorticoid replacement in 92.6%, thyroid replacement in 40.7%, and testosterone in 7.4% of patients. All patients remained on glucocorticoid therapy indefinitely after hypophysitis was diagnosed. ICIs were momentarily held in 22.2% and discontinued in 51.86% of patients, while 25.9% of patients did not experience treatment interruptions. Conclusions: Immune-related hypophysitis is an uncommon irAE that presents with variable clinical and diagnostic findings. Nonetheless it is a cause of permanent glucocorticoid dependence. Healthcare professionals should be aware of this entity so treatment can be promptly started.
Safety and tolerability of HLX07 combined with chemotherapy in patients with advanced solid tumors.
Authors: Li Qun , Guo Ye , Xue Junli , Ge Xiaoxiao , Lin Fengjuan , Xue Liqiong , Zhao Wei , Tang Wenbo , Hu Chen , Ni Hongyan , Li Jing , Wang Qingyu , Zhu Jun , Li Jin
Published: June 2023

e15001

Background: HLX07 is a novel humanized anti-EGFR monoclonal antibody. This study aimed to evaluate the safety, tolerability, pharmacokinetics, and preliminary efficacy of HLX07 combined with different chemotherapy regimens (chemo-regimens) in patients with advanced solid tumors. Methods: This was a single-center, open-label, dose-escalation phase 1b/2 study. Three fixed-dose chemo-regimens were given together with HLX07: 1) gemcitabine 1000 mg/m2 (on Days 1 and 8) and cisplatin 75 mg/m2 (on Day 1) in 3-week cycles for 4–6 cycles; 2) paclitaxel 80 mg/m2 and carboplatin AUC = 2 (both on Days 1, 8, and 15) in 3-week cycles for 4–6 cycles; and 3) mFOLFOX6 (on Day 1) in 2-week cycles for 6–12 cycles. Patients with metastatic or recurrent advanced solid tumors suitable to be treated with the above chemo-regimens were given HLX07 IV at escalating doses (400, 600, or 800 mg, QW) combined with chemotherapies following BOIN design. The primary endpoints were dose-limiting toxicities (DLTs) and maximum tolerated dose (MTD). Secondary endpoints included safety and preliminary efficacy. Results: As of August 28, 2022, 56 patients were enrolled; 21 received HLX07 with gemcitabine and cisplatin, 21 received HLX07 with paclitaxel and carboplatin (n = 3 for HLX07 400 and 600 mg, n = 15 for HLX07 800 mg for each chemo-regimen), and 14 received HLX07 with mFOLFOX6 (n = 3 for HLX07 400 and 600 mg, n = 8 for HLX07 800 mg). No DLT related to HLX07 was reported. The MTD was not reached. A summary of treatment-emergent adverse events (TEAEs) in each group is shown in Table 1. All patients experienced TEAEs. Grade ≥3 TEAEs were reported by 17 (81.0%), 20 (95.2%), and 12 (85.7%) patients in each chemo-regimen group, respectively. Only 1 (33.3%) patient in the HLX07 400 mg plus paclitaxel and carboplatin group experienced TEAE leading to death (upper gastrointestinal hemorrhage, possibly unrelated to study drugs). Conclusions: HLX07 was safe and well tolerated when combined with chemotherapy in patients with advance solid tumors. Clinical trial information: NCT03577704 . [Table: see text]
Pan-cancer analysis to identify molecules interacting with CasL-Like 2 to promote angiogenesis via mTOR/HIF1A/VEGFA pathway in hypoxia tumor environment.
Authors: Song Yu-xin , Li Lei , Lan Jian-yun , Tang Jinhai
Published: June 2023

e15002

Background: Cytoskeleton dynamics generate power for diverse motile and morphogenetic processes in eukaryotic cells. Aberrant actin dynamics are closely associated with pathological conditions and implicated in the occurrence of various tumors. The underlying mechanisms remain to be explored. This study aims to identify a unique redox enzyme, Molecules Interacting with CasL-Like 2 (MICALL2), as a regulator of actin filaments during tumorigenesis and to evaluate its potential value in predicting drug sensitivity to anti-angiogenesis agents. Methods: First, bioinformatic analysis was used to demonstrate MICALL2 expression level across distinct cancers and its relationship with clinical characteristics from TCGA and CPTAC databases. Functional enrichment analysis and whole-mountin situ hybridization were performed. Gene correlation analysis and IHC were used to detect the correlation between MICALL2 and HIF family. Next, chosen kidney renal clear cell carcinoma (KIRC) as a research object, the location of MICALL2 and actin was determined by immunofluorescence. IHC on MICALL2, HIF1A, VEGFA, CD31, CD34 of 44 clinical KIRC tissue samples was performed. The role of MICALL2 in proliferation, hypoxia and angiogenesis of KIRC cells was investigated using EDU staining, CCK-8 assay, ROS analysis, flow cytometry, and tubule formation assays. The effect of MICALL2 on the mTOR-related signal pathway was evaluated by western blot. Last, drug sensitivity analysis was obtained from the GDSC, CTRP, and PRISM databases to reveal the relationship between MICALL2 and anti-angiogenesis agents and cytoskeleton-regulating drugs. Results: MICALL2 expression was elevated in most cancers and correlated with poor survival outcomes. Pan-cancer analysis demonstrated that MICALL2 was co-expressed with HIF family members and strongly linked to angiogenesis pathway. Interfering the translation of MICALL2 during zebrafish embryogenesis resulted in defective caudal vein plexus morphogenesis. Additionally, KIRC patients with high MICALL2 expression displayed enhanced HIF1A expression and increased CD34-labeled neovascularization. In KIRC cells, MICALL2 was found co-located with F-actin. MICALL2 overexpressed KIRC cells exhibited more aggressive behaviors, with the tumor microenvironment remodeling to a more hypoxia condition. Up-regulation of MICALL2 promoted tumor angiogenesis via activating a novel mTOR/HIF1A/VEGFA pathway. Furthermore, tumor patients with high MICALL2 expression were more likely to be resistant to anti-angiogenesis agents and microtubule depolymerization inhibitors, but sensitive to actin polymerization inhibitors. Conclusions: Our results suggest that MICALL2 plays a crucial role in tumor angiogenesis and may be a potential predictor of anti-angiogenic agent sensitivity across multiple cancers.
Population pharmacokinetic and exposure-response safety analyses of mecbotamab vedotin (BA3011) in patients with advanced solid tumors.
Authors: Shen Guoxiang George , Briggs Emily , Crass Ryan , Karumanchi Sravan , Boyle William J. , Baker Michael G. , Martin Philippe
Published: June 2023

e15010

Background: Mecbotamab vedotin (BA3011) is antibody-drug conjugate (ADC) consisting of a conditionally active biologic (CAB) anti-AXL humanized (IgG1) monoclonal antibody conjugated to monomethyl auristatin E (MMAE) using a cleavable linker. BA3011 is being developed for the treatment patients with advanced solid tumors including sarcoma with AXL expression. Methods: As of data cutoff date 19 April 2022, BA3011 was administered by intravenous infusion to 67 patients, with doses ranged from 0.3 to 3 mg/kg once every three weeks (Q3W), 1.2 to 1.8 mg/kg once every two weeks (Q2W) and 1.8 mg/kg twice every three weeks (2Q3W). Population pharmacokinetics (popPK) analysis was performed using sequential modeling approach, where a PK model was first developed for BA3011 ADC alone, and then an integrated model was developed for BA3011 ADC and MMAE. The popPK base model for BA3011 ADC and MMAE was used to generate empirical Bayes estimates of PK parameters for patients in Study BA3011-001 and used for the exposure-response (E-R) safety analysis. E-R time-to-event models were developed to characterize the relationship between BA3011 ADC or MMAE exposure and the time to first incidence of adverse events (AEs). Results: A popPK model with two-compartment disposition and parallel linear and nonlinear elimination of BA3011 ADC, formation of MMAE from ADC, and two-compartment disposition and linear elimination of MMAE, adequately described the PK profiles of BA3011 ADC and MMAE in patients based on model evaluation using prediction-corrected visual predictive checks. Key PK parameter estimates are summarized in the table below. In the E-R safety analysis, statistically significant correlations were demonstrated between BA3011 ADC exposure and all safety endpoints explored (TEAE Grade ≥3, neutropenia Grade ≥3, increase in AST or ALT Grade ≥1, anemia Grade ≥2, and peripheral neuropathy Grade ≥1). Correlations were statistically significant between MMAE exposure and TEAE Grade ≥3, neutropenia Grade ≥3, increase in AST or ALT Grade ≥1, but not anemia Grade ≥2 or peripheral neuropathy Grade ≥1. Time-varying Cmax was selected as the most appropriate exposure metric for each model developed, calculated over the past 24 hours, 1 week, or 3 weeks, depending on the model. Conclusions: The popPK relationship between ADC and MMAE were adequately characterized using a sequential modeling approach. BA3011 ADC exposure was correlated with all safety endpoints evaluated whereas MMAE exposure did not demonstrate correlations with anemia or peripheral neuropathy. These models are essential to generate predictions of exposure and safety for dosing regimens to be evaluated in a planned Phase 2 clinical trial. Clinical trial information: NCT03425279 . [Table: see text]
Tolerability and toxicity profiles of antibody-drug conjugates for the treatment of malignant neoplasms: A meta-analysis of randomized clinical trials.
Authors: Zhou Susu , Kuno Toshiki , Miyagi Etsuko , Wright Jason Dennis , Takagi Hisato , Suzuki Yukio
Published: June 2023

e15011

Background: Antibody-drug conjugates (ADCs) are attractive targeted agents in anticancer treatment due to their unique mechanism of action and reduced toxicity compared to other anti-cancer drugs. Yet little is still known about the adverse events profiles associated with ADCs. Methods: A systematic review was performed including randomized controlled trials (RCTs) of anticancer treatment that evaluated the efficacy of ADCs. PubMed and EMBASE were systematically searched for studies from January 1990 to December 2022. RCTs that were registered in ClinicalTrials.gov and had final reports of comprehensive adverse events were also included. We included studies for patients with both solid and hematologic tumors. The odds ratios (ORs) of treatment-related symptoms, and hematologic, hepatic, ocular, cardiovascular, renal, and respiratory toxicities for patients treated with ADCs and those without ADCs were examined. A random effects model was developed to estimate the pooled ORs of each adverse event associated with ADC use. Subgroup analyses were also performed based on the cancer type (solid tumors versus hematologic malignancy) and ADC regimen (ADC versus ADC plus chemotherapy). Results: 20 studies involving 10,075 patients (5,745 patients with ADC and 4,330 patients without ADC) were included. Among the included trials, 14 (70.0%) were trials for solid tumors and 6 (30.0%) were trials for hematological tumors. Among the included trials, ADC monotherapy was used in 15 (75.0%) experimental arms, and combination therapy of ADC with any other chemotherapy was used in 7 (35.0%). ADCs were associated with a higher risk for all-grade fatigue (OR 1.25, 95% CI 1.08-1.45), anorexia (OR 1.36, 1.09-1.69), nausea (OR 1.46, 1.09-1.97) and sensory neuropathy (OR 2.18, 1.27-3.76). Patients treated with ADCs had a significantly lower risk of all-grade febrile neutropenia (OR 0.46, 0.22-0.96), leukopenia (OR 0.47, 0.29-0.77), and neutropenia (OR 0.56, 0.31-1.01). Whereas, they had a higher risk of all-grade increased alanine aminotransferase (OR 2.51, 1.84-3.40), increased aspartate aminotransferase (OR 2.83, 2.04-3.93), cataract (OR 4.69, 1.86-11.81), blurry vision (OR 3.12, 1.02-9.55), pericardial effusion (OR 4.27, 2.71-6.72), and epistaxis (OR 2.45, 1.50-4.01). In the subgroup analysis, a similar toxicity profile was observed when comparing the solid tumors versus hematologic malignancy groups and the ADC versus ADC plus chemotherapy groups, except for hematologic adverse events, which showed an increased risk in patients with solid tumors and those treated with both chemotherapy and ADCs. Conclusions: This comprehensive profile of adverse events associated with ADC-based treatment shows increased treatment-related symptoms, hepatic, ocular, and cardiovascular toxicities although no increase of high-grade are seen.
Preclinical antitumor activity of an anti-HER2/Trop-2 bispecific antibody-drug conjugate with a new DNA topoisomerase I inhibitor.
Authors: Zhong Helen , Xiong Yan , Huang Han , Pan Yuming , Cao Lijin , Liu Shen , Wang Na , Sun Bing , Liu Dingguo , Yuan Wei , Fang Jan , Bao Haifeng
Published: June 2023

e15013

Background: In recent years, antibody-drug conjugates (ADCs) have made significant progress in cancer therapy and show a promising future. Among the approved ADCs, Enhertu, the HER2 directed ADC and Trodelvy, the Trop-2 directed ADC, have shown considerable clinical benefits. Bispecific antibody-drug conjugate represents a fast-growing class of next generation ADC due to its potentials in enhancing specificity, improving efficacy, and reducing toxicity. Additionally, targeting more than one molecule can be useful to avoid resistance to treatment and broaden the patient population. Methods: We generated a novel bispecific ADC (BIO-201) co-targeting HER2 and Trop-2 which are the two clinically validated tumor-associated antigens that are expressed on a wide variety of tumors. BIO-201 is consisted of a novel bispecific anti-HER2/Trop-2 antibody conjugated with a potent DNA topoisomerase I inhibitor (TOPO Ii) via a cleavable linker. TOPO Ii is a proprietary payload platform developed in house that exhibits similar characteristics including in vitro and in vivo anti-tumor activities and safety profile to DXd. The bispecific antibody was fully characterized and in vitro and in vivo anti-tumor activity of BIO-201 was assessed using preclinical models. Results: The bispecific antibody showed comparable or increased cell binding and internalization compared with the parental antibodies in various tumor cells that express different surface levels of HER2 and Trop-2. In the in vitro cytotoxicity assay, the bispecific ADC BIO-201 effectively killed cancer cells with IC50 in the sub nM range with BIO-201 being most potent against cancer cells that co-express HER2 and Trop-2. The anti-tumor activity of BIO-201 was further demonstrated in vivo in several tumor xenograft models. Treatment of BIO-201 induced significant tumor regression in HER2 or Trop-2 positive tumors, suggesting broader coverage of tumor types than conventional HER2 and Trop-2 ADCs. BIO-201 also showed similar anti-tumor activity to Herceptin-DXd. Conclusions: Taken together, these findings suggest that anti-HER2/Trop-2 bispecific ADC has potential as an effective therapy for a variety of cancers that co-express HER2/Trop-2 or express either of the targets. BIO-201 may be expected to have a similar clinical activity to Enhertu or Trodelvy but treat more patient sub-populations and indications.
A study of antibody conjugates for the treatment of inflammatory breast cancer.
Authors: Yang Teng , Guo Peng , Wei Qing
Published: June 2023

e15016

Background: Inflammatory breast cancer (inflammatory carcinoma) is a rare metastatic malignant tumor, it accounts for about 2% of all breast cancer cases. Despite its low incidence, inflammatory breast cancer is highly invasive, its five-year survival rate is less than 41%, making its prognosis the worst of all breast cancer subtypes, and effective drugs are urgently needed in clinical practice. Therefore, our study develops novel antibody conjugate drugs that can target overexpressed-ICAM-1 in inflammatory breast cancer cells as an original molecular target precisely. Methods: In this study, flow cytometry was used to perform precise and quantitative target screening of inflammatory breast cancer cell lines (SUM-149, KPL4), the endocytosis efficiency of ADC drugs was verified by flow cytometry efficiency analysis and single-photon confocal analysis, and the cytotoxicity of different drugs and cell lines was explored by CCK8 experiment. A nude mouse model of orthotopic xenograft inflammatory breast cancer was constructed, and the in vivo therapeutic effect of inflammatory breast cancer was observed by intravenous injection of two ICAM-1 antibody conjugates (ICAM-1-MMAE and ICAM-1-DXD), also the ADC drug structure with the best therapeutic effect for inflammatory breast cancer was screened. Results: ICAM-1 molecule in a variety of inflammatory breast cancer cell lines were highly over-expressed, and the antibody internalization rate was 51% at 4h, and after the peak the endocytosis efficiency slowed down into the plateau stage; ICAM1-MMAE and ICAM1-DXD are safer than first-line chemotherapy agents for breast cancer; The tumor volume was significantly reduced after administration into the orthotopic breast cancer xenograft model, which further verified the obvious effect of ADC drugs in vivo. Conclusions: Both ICAM-1-MMAE and ICAM-1-DXD have shown significant anti-inflammatory breast cancer effects in vitro and in vivo, and this study provides potential antibody conjugate drugs for targeted therapy of inflammatory breast cancer, which is expected to help improve the poor prognosis of inflammatory breast cancer.
Relationship between EGFR inhibition in lung adenocarcinoma, cell surface expression of the placental antigen ALPP, and efficacy of ALPP-ADC therapy.
Authors: Katayama Hiroyuki , Chen Yihui , Hong Monica , Xu Hanwen , Vykouka Jody , Park Soyoung , Cai Yining , León-Letelier Ricardo A. , Irajizad Ehsan , Hsiao Fu Chung , Dennison Jennifer , Ostrin Edwin J. , Fahrmann Johannes , Hanash Samir M.
Published: June 2023

e15018

Background: In normal tissues placental alkaline phosphatases (ALPP) and ALPPL2 are exclusively expressed in the placenta. Their discovery in the cancer surfaceome has opened the possibility of targeting ALPP/ALPPL2. However, their low expression in most cancer types limits their therapeutic potential. In this study, we report a novel mechanism of ALPP upregulation in cancer cells that renders these cells more susceptible to ALPP-targeting therapies, which we evidence in the context of lung adenocarcinoma. Methods: Surface expression of ALPP and ALPPL2 was determined following biotinylation tagging of the cancer surfaceome, isolation of biotinylated proteins using monomeric avidin and mass spectrometry of the biotinylated trypsin digested fraction. ALPP antibody was conjugated with Monomethylauristatin F (ALPP-ADC-MMAF). For orthotopic xenograft models of LUAD were injected. For gefitinib treatment, Mice xenotransplanted with H1650 were treated with gefitinib (50mg/kg) daily for 10 days starting from Day 17 post cancer cell inoculation. For the gefitinib/ALPP-ADC combinational treatment, mice received anti-ALPP-MMAF (5mg/kg) at Day 19 and Day 26. Results: We assessed ALPP and ALPPL2 surfaceome protein levels in 158 cancer cell lines and their mRNA expression levels in 10,967 tumors representing 32 cancer types from The Cancer Genome Atlas (TCGA), which revealed ALPP, and to a lesser extent ALPPL2, to be variably expressed in several cancer types including lung adenocarcinoma (LUAD). We further confirmed surface expression of ALPP by tissue microarray analysis of 204 lung tumors. We report that treatment of LUAD cells with EGFR inhibitors or induction of cancer cell quiescence greatly enhanced ALPP surface expression. Mechanistically, repression of EGFR signaling resulted in activation of the transcription factor FoxO3a, which triggered expression of ALPP. Using xenograft models of LUAD, we further demonstrate that gefitinib treatment significantly upregulated surface expression of ALPP in LUAD tumors but not normal tissue. Combination therapy with gefitinib and an ALPP-ADC-MMAF resulted in superior anti-cancer efficacy compared with gefitinib or ALPP-ADC-MMAF alone. Conclusions: Our study provides mechanistic insights into ALPP upregulation in cancer cells and establishes FoxO3a as a transcriptional regulator of ALPP. Importantly, our findings have yielded a novel ‘two-hit’ combinational therapeutic strategy with enhanced ALPP targeting for greater efficacy.
Epigenetic profile of elevated cell free circulating nucleosomes in non-Hodgkin lymphoma.
Authors: Van den Ackerveken Priscilla , Lobbens Alison , Pamart Dorian , Rommelaere Guillaume , Herzog Marielle
Published: June 2023

e15032

Background: Non-Hodgkin lymphoma (NHL) is a common type of hematological malignancy. The non-specific symptoms of lymphoma often delay diagnosis that is mainly based on a lymph node biopsy. However, due to their invasive nature, tissue biopsies have many limitations especially for monitoring therapeutic response where multiple biopsies may be required. Conversely, liquid biopsy offers a promising diagnosis and monitoring tool for cancer detection as it is noninvasive and easily repeatable over time compared to tissue biopsy. In this study, we investigated the circulating levels of intact nucleosomes containing the histone H3.1 isoform (Nu.Q-H3.1) in NHL and healthy subjects. Then, as histone post-translational modifications (PTMs) of circulating nucleosomes are described as potential biomarkers of various solid cancers, we investigated the epigenetic profile of nucleosomes from NHL patients following nucleosome enrichment (Nu.Q Capture) combined with mass spectrometry (MS) compared to healthy donors. Methods: In plasma K2EDTA from 9 NHL and 5 healthy subjects, we measured levels of Nu.Q-H3.1 combined with the epigenetic profile of these circulating nucleosomes using our Nu.Q Capture- mass spectrometry protocol. Subsequently, the capability of the identified histone PTMs to differentiate between NHL and healthy plasma was tested in an independent cohort (n = 24 NHL patients and n = 35 healthy donors) and in two patients undergoing treatment course using available quantitative Nu.Q immunoassays. Results: We describe a high level of circulating H3.1-nucleosomes in human plasma NHL patients compared to healthy subjects. This level of H3.1-nucleosomes was correlated with an elevated concentration of cfDNA and is consistent with the DNA distribution size observed by the Bioanalyzer around 146 bp. In addition, the mass spectrometric analysis of the nucleosome enriched extracts allowed us to profile the epigenetic pattern of circulating nucleosomes in NHL patients. Our analysis demonstrated increased levels of H3.1 as well as the lysine acetylation and methylations of the histone H3 in plasma from NHL patients. Furthermore, the clinical validation of the histone PTMs pattern using immunoassays identified a significant increase in the levels of H3.1-nucleosomes as well as 6 histone PTMs (H3K9Me1, H3K27Me3, H3K36Me3, H3K9Ac, H3K14Ac, H3K18Ac) in NHL patients in comparison with healthy donors. Finally, the application of this histone PTMs pattern on NHL patients demonstrated the potential use of these new biomarkers present in liquid biopsy to evaluate the therapeutic response and monitoring of tumor progression. Conclusions: Our results indicate that levels of Nu.Q-H3.1 are particularly elevated in NHL patients and may be a useful diagnostic tool. Moreover, our work emphasizes the crucial roles of the epigenetic marks present on circulating nucleosomes to detect and monitor diseases such as lymphoma.
Novel blood-based assay for detection of early stage multi-cancer.
Authors: Gregg Jeffrey P. , Michor Franziska , Gilley Caitlin , Williams Miguel , Brown Esther , Gowen Kyle , KashefHaghighi Dorna , Sun James , Kia Amirali , Charlton Jocelyn
Published: June 2023

e15035

Background: Early cancer detection has the potential to significantly improve patient outcomes and reach the Cancer Moonshot goals of reducing the death rate from cancer by at least 50 percent over the next 25 years. Harbinger Health is pioneering early cancer detection with a blood-based test that combines recent genomic and epigenomic discoveries of early cancer and biology-informed artificial intelligence. Unlike prior approaches that are purely statistical to identify informative biomarkers, Harbinger’s approach is informed by insights into specific biological events early during tumorigenesis and is therefore optimized for detecting cancer in patients with very low levels of circulating tumor DNA. Methods: Here, we present data generated using the Harbinger Health assay from 1,046 subjects, 621 with newly diagnosed, treatment-naive cancer (15 different cancer types) and 425 individuals with no history, diagnosis, or cancer symptoms. Using this sample cohort, we developed and applied a rigorous framework for training, calibration and predicting likelihood of cancer using a multi-layered logistic regression-based machine learning algorithm, generating final outputs of binary classification (cancer yes/no). Our framework involved multiple iterations of 10-fold cross-validation of the full dataset, and we report solely on samples that appear in the held-out test set in each iteration. Additionally, we developed a similar framework to predict tissue of origin (TOO) for cancer samples. Results: The overall sensitivity of cancer detection was 82% (95% confidence interval (CI): 72.7-91.0%) at 95% specificity. Notably, the sensitivity was 74% (95% CI: 54.8-92.7%) for stage 1 and 84% (95% CI: 65.3-100%) for stage 2. Furthermore, we were able to correctly predict cancer in 95% of patient samples with at least 0.037% tumor fraction. The overall sensitivity for high incident cancers were: breast (73%), prostate (82%), lung (85%), and colorectal (96%) at 95% specificity and the overall accuracy of TOO prediction was 86% when the tumor fraction was greater than 0.1% in the top 3 most prevalent cancer types (breast, colorectal and lung). Importantly, we noted that technical variability was introduced when performing assay optimization strategies, yet we observed comparable performance when assessing subsets of stably processed samples, indicating that our performance is robust. Conclusions: Overall, these results demonstrate that the Harbinger Health platform, with its biology-informed approach, has the potential for highly sensitive multi-cancer diagnostic accuracy and specifically those with early-stage cancer. The platform is now being validated in a 10,000-subject prospective clinical trial (CORE-HH/NCT05435066).
Utility of next generation sequencing (NGS) in advanced malignancies in a community hospital in an underserved area: A game changer?
Authors: Desai Krishna , Iqbal Sabah , Baralo Bohdan , Pereira Kristal , Jain Akhil , Deshpande Sohiel , Renzu Mahvish , Mehta Vidhi , Rather Manzoor , Thirumaran Rajesh
Published: June 2023

e15037

Background: Next Generation Sequencing (NGS) is a revolutionary test used in advanced malignancies to study specific mutations, define tumor burdens, & design personalized treatment plans based on the target mutations. With this study, our aim is to highlight the role of NGS, especially in a community hospital & emphasize on its implementation as a standard of care in advanced metastatic cancers. Methods: We retrospectively reviewed the charts of 51 patients after IRB approval, aged > 18 years with no specific demographic restrictions, with advanced malignancies who had NGS at Mercy Catholic Medical Center, PA. Results: Of 51 patients, 25 were males & 26 were females, 44 patients had positive results on NGS. 100 types of mutations were identified & 338 mutations in total were found. Lung cancer had highest mutations (41.17%-21/51), followed by colon (25.49% - 13/51), breast, ovarian & endometrium (13.72% 7/51), & the rest comprised of GI malignancies & prostate cancers. Of 100 mutations, 32% (32/100) had actionable targets & received NGS directed chemoimmunotherapy, 22% (22/100) mutations had experimental chemo-immunotherapeutic drugs, & 49% (49/100) had no specific NGS directed therapy actionable/target mutation. The most common (1,2,3) & some rare mutations (4,5 and 6) with directed immunotherapy noted on NGS are listed in Table 1. Conclusions: As per our study, 86.27% of the sample size showed positive results on NGS. 32% of these mutations had NGS directed chemoimmunotherapy against these target mutations. In contrary to standard treatment regimens, a personalized treatment plan was developed for these patients, thereby curbing further progression & improving survival in patients with advanced cancer. For example, a patient with stage IV colon cancer who was progressing on the standard treatment with FOLFOX (Folinic acid, flurouracil and oxaliplatin), was started on Ivosedenib, after undergoing NGS and showed marked improvement clinically and radiologically. 29 out of 43 guidelines for various malignancies established by National Comprehensive Cancer Network highlight the significance of NGS. With this study, our goal is to emphasize on the utility of NGS in patients with advanced cancer & the need for it to be incorporated as the standard of care, in small & mid-tier community hospitals, which would need a devised plan for its implementation. [Table: see text]
Comprehensive multi-omic testing from a single laboratory to provide information in ovarian cancer.
Authors: Mancini-Dinardo Debora , Lenz Lauren , Comeaux Matthew , Hatcher Shanell , Gutin Alexander , Thomsen Anne , Wroblewski Ashley , Eich Alexzandra , Moll Tyson , Poyser Lynette , Slavin Thomas Paul , Timms Kirsten ,   Precise Treatment Steering Committee
Published: June 2023

e15045

Background: Molecular profiling of ovarian tumors allows for personalized disease treatment but often requires testing from multiple laboratories. This study highlights results from a molecular testing pathway that reports on germline and tumor mutations, microsatellite instability (MSI), tumor mutation burden (TMB), and homologous recombination deficiency (HRD) to guide optimal treatment selection. Methods: Samples from a consecutive prospective cohort of ovarian cancer patients were collected and tested with an HRD companion diagnostic (CDx) test (FDA-approved) that assesses genomic instability score (GIS) via a 3-biomarker signature and tumor mutation screening using a 523-gene DNA and a 55-gene RNA panel to identify clinically actionable test results using Association for Molecular Pathology (AMP) tiering. Some patients also received germline analysis via a 48-gene panel or a BRCA1/2 CDx. Clinical data (tumor stage and grade, histological subtype) were compared using logistic regression models. GIS distributions were compared using Kolmogorov-Smirnov tests. Results: To date, results from 389 eligible patients have been analyzed. Table 1 shows the primary results of molecular testing. Tumor subtype analyses revealed that high-grade serous subtype was tested most frequently (66.1%). Some less-represented subtypes (e.g., carcinoscarcoma, mucinous) appeared to have GIS distributions like high-grade serous, while others generally have low GIS. GIS distributions were significantly different in high-grade (vs moderate and vs low-grade) and stage I (vs the similar stages II-IV) tumors. Tier 1A/B mutations were observed in 16.7% of tumors; 36.9% of those were non- BRCA pathogenic variants/mutations. Distributions of histological subtype and grade were significantly different between t BRCA+ and non- BRCA samples with other Tier 1A/B mutations. Mutations were observed in 9.0% of germline tests; tumor mutation testing identified all germline variants except for a PMS2 multi-exonic deletion. Conclusions: Multi-omic testing found ~30% of ovarian cancer patients received a clinically actionable test result (HRD was the most common) and ~90% of results conferred eligibility for clinical trials. Incorporation of germline testing and RNA analysis identified unique actionable findings. A single source for comprehensive testing combined with a single integrated report facilitates rapid testing, test interpretation, and treatment choice. [Table: see text]
Landscape of FGFR2/3 alterations in genitourinary cancer.
Authors: Che Xiangyu , Liu Huanhuan , Qin Jiayue , Cao Shanbo
Published: June 2023

e15047

Background: FGFR2/3 is frequently activated by mutation or overexpression, and it is a validated therapeutic target in several cancers. However, the molecular profile of FGFR2/3 in genitourinary cancer is still lacking. In this study, we assessed FGFR2/3 alterations for better understanding therapeutic implications in genitourinary cancer. Methods: Patients diagnosed with genitourinary cancer were enrolled in the study. Tumors and plasma samples of participants were sequencing for next-generation sequencing from Chinese cohort and TCGA database. Results: A total of 3858 patients were enrolled with 1640 renal cell cancer (RCC), 1300 prostate cancer (PC), and 918 urothelial cancer (UC), containing 2482 tumor tissues and 1376 baseline plasma samples were collected. A total of 14.6% patients (563/3858) harbored at least one FGFR2/3 alterations . 24.1% (221/918) UC carried the most FGFR2/3 alterations , including somatic mutation (18.2%), CNV (5.7%), and fusion (2.1%). Furthermore, the altered FGFRs targets are mainly S249C (6.5%), Y373C (2.5%), FGFR3-TACC3 fusion (1.6%). R248C (1.2%), and G370C (0.5%) in UC. FGFR2/3 alterations were found in 14.0% (182/1300) PC, including somatic mutation (8.2%), CNV (5.8%), and fusion (0.4%). However, only 2 patients were found to have altered FGFRs targets including R248C mutation and FGFR3-TACC3 fusion (n = 1 each). Regarding of RCC, only 9.8% (160/1640) patients had FGFR3 alterations , including somatic mutation (5.8%), CNV (5.0%), and fusion (0.5%). Furthermore, the altered FGFRs targets are mainly R248C, S249C, Y373C (n = 4 each). Moreover, patients with FGFR2/3 alterations harbored a significantly higher TMB than those in patients without FGFR2/3 alterations (Mean: 13.3 vs 9.4; p < 0.001). There was a significant difference in TMB among different cancers with FGFR2/3 alterations , with UC being the highest and PC the lowest (Mean: 17.0 vs 13.0 vs 10.2; p < 0.001). Conclusions: FGFR2/3 alterations are frequently altered in genitourinary cancer, especially UC. Comprehensive genomic profiling of FGFR2/3 alterations highlights the clinical and therapeutic implication.
Utilize protein markers and genomic features for minimal residual disease detection in hepatocellular carcinoma.
Authors: Tu Shichun , Li Shiyong , Ren Qingqi , Zhong Guolin , Li Yunfei , Feng Yumin , Zhang Yixia , Chang Yinyin , Long Guanghui , Zhu Dandan , Geng Shuaipeng , Mao Mao
Published: June 2023

e15054

Background: Several studies have shown the clinical value of using circulating tumor DNA (ctDNA) for minimal residual disease (MRD) detection, which is more sensitive than traditional imaging methods. However, most MRD detection technologies focus on identifying mutations and require cancer tissue samples. These methods often have limitations, such as high cost, inaccessible cancer tissue, or inability to overcome tumor heterogeneity. SeekInCure is a cost-effective multi-dimensional MRD test that does not require cancer tissue analysis. The purpose of this prospective study was to evaluate the potential broad clinical utility of SeekInCure for detecting MRD in patients with hepatocellular carcinoma (HCC). Methods: 34 HCC patients undergoing radical surgery were prospectively enrolled in this study from Peking University Shenzhen Hospital. Peripheral blood samples of 8 mL were collected before and after surgery and analyzed using the SeekInCure assay, which integrates the protein tumor marker (AFP) and cancer genomic hallmarks: copy number aberrations and fragment size from ctDNA. Patients with positive results for AFP and/or ctDNA in the preoperative/postoperative samples were defined as MRD-positive (MRD+), while negative results were defined as MRD-negative (MRD-). Results: Of the 34 patients, 73.5% (25/34) were in early stages (stages I/II: 61.8%/11.8%), and 61.8% (21/34) received adjuvant therapies. The median follow-up was 279.5 days, ranging from 90 to 1,253 days. 82% (28/34) of the patients had MRD+ results in the preoperative samples, which showed remarkable sensitivity in a cohort with more than 70% of early-stage patients. Our previous findings were confirmed, with preoperative MRD+ patients having worse survival than MRD- patients1. After radical surgery, 26.5% (9/34) were MRD+. MRD+ patients had worse survival than MRD- patients, with a median overall survival (OS) time of 298 days for MRD+ patients, while the OS time for MRD- patients was not reached (P < 0.01). 6 patients with MRD- in both pre- and post-operative samples had a very favorable outcome (100% OS). Conversely, 9 patients with both positive results had the worst outcome. We also found that MRD- patients receiving adjuvant therapies did not have a survival benefit compared to those without adjuvant therapies, but MRD+ patients receiving adjuvant therapies had better OS than those without adjuvant therapies (P = 0.026). Conclusions: This study confirms the excellent performance of our SeekInCure assay, which does not require cancer tissue analysis, in detecting MRD and the prognostic value of MRD in HCC patients. Adjuvant has no influence on survival, especially in MRD- patients, which indicates that amost one-third of HCC patients do not need additional adjuvant therapy.