bluuebunny/crossref_metadata_2025 huggingface dataset:
Homologous recombination-related (HRR) gene mutations in patients with glioma and their relevance to genomic characteristics, tumor mutation burden (TMB), and prognosis.
Authors: Zheng Zhiming , Li Mengmeng , Zhang Qin , Luo Ningning , Qi Yingxue , Sun Tingting , Qi Chuang
Published: June 2023

e14023

Background: Homologous Recombination Repair (HRR) is one of the core repair methods for DNA double strand damage and the mutations of HRR pathway gene may be the main cause of many cancers. However, there is little research on the potential correlation between the molecular characteristics of HRR pathway genes and glioma. Our study explored HRR gene mutations in glioma patients and its relevance to genomic characteristics, tumor mutation burden (TMB) and prognosis. Methods: Samples from 367 glioma patients with a median age of 46 were detected by 551-gene panel or whole-exome sequencing (WES), and the WES data of 894 glioma patients were extracted from TCGA database ( http://cancergenome.nih.gov ). Among 367 glioma patients, 151 patients occurred HRR somatic mutations. Among them, 47.68% (72/151) patients carried HRR related pathogenic mutation (P/LP), and 52.32% (79/151) patients carried HRR benign/likely benign/variant of uncertain significance mutations (B/LB/VUS). The patients were divided into two groups according to whether patients carried HRR related pathogenic mutation, HRR P/LP group (72) and Non-HRR P/LP group (295). The differences of genomic characteristics, TMB and prognosis between HRR P/LP group and Non-HRR P/LP group were analyzed. Results: In 151 glioma patients harboring HRR somatic mutations, PTEN had the highest mutation frequency (49%; 74/151), followed by CHEK2, DNMT3A, BRCA2 and ATM, with mutation frequency of 11% (17/151), 10% (15/151), 9% (14/151）and 9% (14/151). Patients of HRR P/LP group and HRR B/LB/UVS group showed a significant difference at HRR pathway genes, including PTEN, RAD50, MSH6 and RAD21. In addition, differences of non-HRR pathway genes in HRR P/LP group and Non-HRR P/LP group were found. The mutation frequency of these differential genes was generally high in HRR P/LP group. However, compared to Non-HRR P/LP group, the mutation frequency of IDH1 in HRR P/LP group was significantly higher (Non-HRR P/LP 32% VS HRR P/LP 10%; P＜0.0001). The TMB value in Non-HRR P/LP group was significantly higher than that in HRR P/LP group (P＜0.0001), and the same trend of TMB in two groups was discovered in TCGA cohort (P＜0.0001). Importantly, a significant difference of overall survival (OS) in TCGA database was found between HRR P/LP group and Non-HRR P/LP group (P＜0.0001, HR =2.17, 95%CI: 1.54-3.06), and the median OS were 17.6 and 34.9 months, respectively. Conclusions: This study showed HRR pathway genes status was significantly related to the genomic characteristics, TMB and prognosis of glioma patients. Compared to patients in HRR P/LP group, patients in Non-HRR P/LP group had higher mutation frequency of IDH1, lower TMB value and better prognosis. The relationship between HRR pathway genes and glioma needs to be further explored.
Results from the first step of a phase II study of vaccination with autologous dendritic cells loaded with autologous tumour homogenate in glioblastoma.
Authors: Ridolfi Laura , Gurrieri Lorena , Riva Nada , Bulgarelli Jenny , De Rosa Francesco , Guidoboni Massimo , Fausti Valentina , Ranallo Nicoletta , Tazzari Marcella , Petrini Massimiliano , Granato Anna Maria , Pancisi Elena , Foca Flavia , Dall'Agata Monia , Amadori Elena , Cortesi Pietro , Gamboni Alessandro , Pasini Giuseppe , Ibrahim Toni , Tosatto Luigino
Published: June 2023

e14025

Background: Glioblastoma (GBM) is a poor prognosis malignant grade IV glioma. After surgical resection, standard therapy consists of concomitant radiotherapy (RT) and temozolomide (TMZ) followed by TMZ alone. Multiple phase I/II trials and at least 3 meta-analysis showed improved survival (OS) and progression free survival (PFS) with DC vaccination in High-grade gliomas (HGGs) patients (pts). In pts developing antitumor immunity, DC vaccination increases the amount of intratumoral activated cytotoxic T lymphocytes and decreases the number of FoxP3 positive regulatory T cells. Based on these data we have developed a phase II protocol with DC vaccine concomitant to standard RT-CT in pts who have undergone radical surgery for GBM. Methods: This is a single-arm, monocentric, phase II trial of a DC vaccination integrated to standard therapy in resected GBM pts. All pts receive a dendritic cell vaccine loaded with autologous tumor homogenate for up to one year. The vaccine administration starts at the end of the RT-CT (Induction Phase) and then is alternated to TMZ cycles (Maintenance Phase). Primary end points are PFS and safety, among secondary end points there are the in vitro (Elispot, Plasma Cytokines, Tumor tissue analysis) and in vivo (DTH skin test) Immune response evaluations. A Simon's two-stage design has been used for the sample size calculation. In the first stage, 9 patients have been accrued and a total of 28 patients will be enrolled at the end of the second step. Results: From July 2021, the 9 pts of the first step has been enrolled, 4 females and 5 males with median age of 58 years. Five pts had no MGMT mutation. All 9 pts concluded the induction phase. To November 2022 four pts have progressed with a median PFS, from the date of leukapheresis, of 6.5 months (3.2-NE). DTH test became positive in 5 out of 9 evaluable pts. Median OS from the date of surgery was 11 months. No gr 3-4 vaccine related toxicities have been observed and gr 1-2 toxicities were mostly due to local skin reactions. Conclusions: On the basis of the results described above and the vaccine favorable toxicity profile the Study Team decided to proceed with the second step. Clinical trial information: EUCTR220-003755-15 . [Table: see text]
The activity and safety of anlotinib for patients with recurrent malignant glioma: A single-center, retrospective study.
Authors: Sun Xudong , Li Min-Jie , Zhang Xue-ying , Shen Yue-feng
Published: June 2023

e14032

Background: Anlotinib is a novel tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor (VEGFR) 1/2/3, platelet-derived growth factor receptor (PDGFR), fibroblast growth factor receptors (FGFR) 1/2/3/4, c-Kit, and Ret. We report results from this retrospective study to determine the efficacy and tolerability of Anlotinib as a treatment for recurrent malignant glioma (rMG). Methods: A total of 26 eligible patients who relapsed from the standard chemoradiotherapy regimen (TMZ and radiotherapy) after surgery because of tumor located in the eloquent brain areas were enrolled in this study between May 2020 and August 2022. Patients were subjected to a concurrent treatment of Anlotinib (12mg qd) until disease progression or intolerable toxicity. Efficacy was evaluated using Response Assessment in Neuro-Oncology criteria for high-grade glioma. Safety was assessed using NCI-CTCAE 5.0. Survival was estimated with the Kaplan-Meier curve and log-rank test. Results: In total, 26 (11 male, 15 female) patients with recurrent malignant glioma were enrolled. The disease control rate was 84.6% (95%CI 36.9%-76.7%) and the objective response rate was 57.7%(95%CI 65.1%-95.6%). The median progress-free survival (PFS) for all patients was 9.0 months (95%CI 2.88-15.19). The most common treatment-related adverse events were hand-foot syndrome (38.5%), Weakness (38.5%), hypertension 30.8%) and there were no Grade ≥3 toxicities. Conclusions: The present study showed that anlotinib had a promising efficacy and favorable tolerability in treat rMG. Further randomized controlled clinical studies are needed to confirm the efficacy of Anlotinib for the treatment of rMG.
Genomic landscape of CDK6 amplification and its potential prognostic value in adult-type diffuse gliomas.
Authors: Yuan Xi , Que Tianshi , Yi Guozhong , Li Zhiyong , Zheng Haojie , Liu Yawei , Wang Xiaoxuan , Zhang Xing , Huang Guanglong
Published: June 2023

e14038

Background: Glioma is commonly described as a disease of neuroglial cell proliferation. And the gene related to cell cycle pathway alter frequently in glioma including CDKN2A/B, CDK4 and CDK6. CDKN2A/B homozygous deletion is proved to be associated with worse survival in IDH-mut gliomas and meningiomas. However, the correlation between CDK4/6 amplification and the prognosis of glioma remains unclear. Herein, we explored the distribution of CDK4/6 alterations and whether they are the prognostic biomarker of glioma. Methods: A total of 90 patients with adult-type diffuse glioma were enrolled to this retrospective study from our center. Whole genome sequencing (WES) was performed to identify the mutations of the cohort. Results: CDK4 and CDK6 amplified in 17 (18.9%) and 12 (13.3%) patients, respectively. Four patients harbored both CDK4 and CDK6 amplification. The Cox multivariate regression revealed only CDK6 amplification was the independent factor (HR 4.0, 95% CI 1.1-15.2, p=0.041) of overall survival (OS). Interestingly, CDK6 amplification occurred easier in female and trended to be co-mutant with CDKN2A/B homozygous deletion. CDK6 amplification was found to be significantly associated with shorter OS ( p<0.05) of glioblastoma (GBM). Although TERTp was not related to the OS of GBM according to TCGA, patients harboring both TERTp and CDK6 amplification had the significant worst OS ( p<0.001). Conclusions: Our study compared the mutational characteristics of gliomas with CDK4/6 alterations and demonstrated the potential prognostic value of CDK6 amplification, and provide the new basis of precise diagnosis and treatment in gliomas.
Anlotinib for patients with newly diagnosed glioblastoma with unmethylated MGMT promoter: An open-label, single-center, phase 2 clinical trial.
Authors: Yang Kunyu , Wu Bian , Zhang Zhanjie , Peng Gang , Huang Jing , Hong Xiaohua , Ding Qian , Shi Liangliang , Wang Xuan , Zhao Hongyang , Jiang Xiaobing
Published: June 2023

e14039

Background: Glioblastoma (GBM) is the most common primary malignant brain tumor and one of the most deadly forms of cancer in humans. Given the temozolomide’s minimal benefit in O6-methylguanine-DNA Methyltransferase (MGMT) unmethylated GBM, new and effective therapies are urgently needed. In this study, we aimed to assess the safety and efficacy of anlotinib, a novel multitarget tyrosine kinase inhibitor for VEGFR, PDGFR, and c-Kit, in patients with MGMT unmethylated GBM. Methods: This was an open-label, single -center, single-arm, phase 2 trial. Eligible patients included those aged 18 to 70 years who had newly diagnosed histologically confirmed GBM with unmethylated MGMT promoter region. The cutoff value ≥ 10% was used to classify MGMT methylated vs. unmethylated cases. After initial surgical resection, patients with GBM received concurrent radiotherapy (2.0 grays [Gy]/day; total dose, 60 Gy) and anlotinib (beginning on day 43 after surgery, at a dose of 12 mg orally once daily on days 1-14 of a 21-day cycle), followed by 6 cycles of maintenance therapy with anlotinib (at a dose of 12 mg orally once daily on days 1-14 of a 21-day cycle). Patients were re-evaluated every six weeks. The primary endpoint was progression-free survival (PFS), which is defined as the interval from the initiation of anlotinib treatment until the date of disease progression or death. The secondary endpoints were overall survival (OS) and safety. Standard Response Assessment in Neuro-Oncology (RANO) criteria were used to determine tumor response and progression. Adverse events were assessed according to the National Cancer Institute Common Terminology Criteria for Adverse Events, version 4.0. This trial was registered with chictr.org.cn, ChiCTR2000040116. Results: A total of 32 participants were enrolled from September 2020 to June 2022. All patients were with IDH wild-type and MGMT unmethylated status. The median age was 50 years (range, 27-65), and 19 (59.4%) were male. The median follow-up duration was 14.8 months (range, 5.5-23.7). The median PFS was 10.5 months (95%CI: 8.5-12.5), and the median OS was not reached. Treatment-related adverse events (TRAEs) occurred in 13 (40.6%) patients of patients and grade 3 adverse events were observed in 4 (12.5%) patients. The most common toxicities were hypertension (13 [41%]), hand-foot syndrome (12 [38%]), hematuresis (9 [28%]) and liver dysfunction (5 [16%]). Grade 3 therapy-related adverse events included liver dysfunction (2 [6.3%]), hypertension (1 [3.1%]), hand-foot syndrome (1 [3.1%]). No grade 4-5 or unexpected adverse events were observed. Conclusions: This preliminary analysis suggested that anlotinib had promising efficacy and favorable tolerance as treatment of newly diagnosed GBM with unmethylated MGMT promoter. Clinical trial information: ChiCTR2000040116 .
Mutation profiling of gliomas based on sex, age, and family history.
Authors: Fan Xue , Yang Ming , Zhang Xiaoni , Li Hui , Wan Xuebin , Liu Yuanyuan , Zhou Shuxiong
Published: June 2023

e14041

Background: The gender, age and family history of patients are important factors leading to the mutation of tumor-related genes. Currently, the relationship between these factors and tumor gene mutations in glioma is unclear. Therefore, we identified the effect of gender, age, and family history on cancer-susceptibility gene mutations in patients with glioma through retrospective analysis. Methods: A total of 735 FFPE tissue samples were collected from 2019 to 2022 in glioma patients, and sequenced by HapOnco StarPanel NGS Assay (covering 1326 genes + 3 viral sequences). Results: (1) These mutations were categorized into four subgroups according to the age of the patients. In the 0-20 years group, the main mutated genes were TP53 (63.64%), BRAF (36.36%), and H3F3A (22.73%) in females (n = 22) and TP53 (26.92%), BRAF (46.15%) and H3F3A (19.23%) in males (n = 26), respectively. In the 21-40 age group, the gene mutation rates of TP53, IDH1, and TERT were 71.43%, 58.57%, and 21.43% in female (n = 70), whereas these genes ( TP53, 72.90%; IDH1, 63.55%; TERT, 35.51%) displayed higher mutation frequency in males (n = 107). In the 41-60 age group, TERT (61.70%), TP53 (43.97%), and IDH1 (36.88%) in females (n = 141) showed obvious difference compared with that ( TERT, 54.33%; TP53, 47.12%; IDH1, 42.31%) in male (n = 208). Aged patients, especially those over 61 years, the most mutated genes were TERT (63.08%), EGFR (60.00%), and TP53 (49.23%) in female (n = 65), respectively. However, the most commonly mutated genes in male presented apparently different than the genes in female such as LRP1B, CSMD3, SYNE1, NF1, ATM, PTEN, RB1, TERT, MET, and TP53. (2) Generally, the peak incidence of patients with glioma is between 41-60 years old. We found the percentage of the total incidence of glioma patients was associated with the family history (positive, 56.25%; negative, 47.68%). Conclusions: The gene mutated rates were depending on the age, sex, and family history in glioma, which might reveal potential effect for biomarker detection in combination with these categories in clinical.
Concurrent chemoradiation and adjuvant chemotherapy in newly diagnosed glioblastoma multiforme: A retrospective analysis of single center.
Authors: Iruarrizaga Eluska , Iza Estibaliz , Mañe Joan Manel , Rubio Itziar , Novo Eneko , Lombardero Lara , De Miguel María José , Muñoz Alberto
Published: June 2023

e14053

Background: The standard of care of glioblastoma multiforme (GBM) first line therapy is maximum safe debulking and posterior treatment based in concurrent chemo-radiation (Qt-Rt) followed by temozolomide-based chemotherapy. Despite a complete resection and adjuvant treatment, overall survival (OS) in patients (pts) with GBM is poor and it depends on tumor biology and patient characteristics. We analyzed retrospectively clinical and tumor biology characteristics of pts with GBM treated in our center with Qt-Rt +/- adjuvant Qt and their impact in OS. Methods: From November-2016 to October-2022, we collected data for 82 pts (median age: 60 years) who underwent Qt-Rt exclusively (25 patients) or Qt-Rt and adjuvant temozolomide (57 patients). Pts characteristics are detailed. Results: In multivariate analysis, significant difference of OS were observed in function of type of surgery (better OS in R0; p=0.015), performance status (better OS in PS 0-1; P=0.006), MGMT methylation (better OS in methylated; p=0.037), ATRX status (better OS in negative; p=0.05). No differences were founded in function of TERT (p=0.47) or p53 status (p=0.57). OS was significantly increased in pts treated with Qt-Rt and adjuvant Qt (17.7 +/- 1.7 months) vs Qt-Rt exclusively (11.35 +/-1.62 months) (p=0.013). After Qt-Rt treatment, radiological response was (%): no evidence of disease 13.4/partial response 6.1/stable disease 58.5/progressive disease 18.3/not evaluated 2.4. Most relevant adverse event G3/4 during Qt-Rt (%): thrombocytopenia 7.31, neutropenia 8, leukopenia 3.65, anemia 2.4, thrombosis 2.4. There were 2 toxic death (infection grade 5). After Qt-Rt, 57 pts received adjuvant treatment (median number of cycles: 5; range 1-15). Most relevant toxicity G3/4 during this therapy was (%): asthenia 3.05, anorexia 1.75, thrombocytopenia 5.26, neutropenia 3.05, leukopenia 1.75. There was 1 toxic death (infection G5). Pts that received 4 or more cycles of adjuvant temozolomide had a significant impact on OS versus those that completed 3 or less cycles (19.28 +/- 2 months vs. 12.05 +/- 1.5 months; p=0.022). Conclusions: Temozolomide-based chemotherapy in GBM is an effective and safe therapy and the premature interruption of this treatment may impact on OS. It is essential to know patient characteristics and tumor biology in order to identify patients that will beneficiate from this treatment.[Table: see text]
Pre-clinical efficacy of the first glyco-humanized oncolytic polyclonal antibody (XON7) in onco-hematology.
Authors: Ciron Carine , Shneiker Françoise , Duvaux Odile
Published: June 2023

e14519

Background: Polyclonal antibodies pAb were shown to possess oncolytic properties a century ago with reported clinical responses. More recent pre-clinical models confirmed pAb efficacy though their ability to tackle complex target antigens. The emergence of escape variants could be minimized by multi-targeting of tumor cells with pAb, which would be more efficient than mAb at mediating effector functions for target destruction. However, the use of heterologous pAb is limited by their immunogenicity. Glyco-humanization (GH) solves this issue and allows to reconsider the potential of pAb in oncology. 3 GH-pAb have already been introduced in clinic in non-oncology settings, with a good safety profile. XON7 is a new GH-pAb targeting cancer cells. Methods: Complement dependent cytotoxicity (CDC) and apoptosis against various tumor cell lines and PBMC from healthy donors were tested with XON7 at increasing concentrations (6,25 µg/mL to 400 µg/mL) Specific binding to human tumors was assessed by immunochemistry on tissue micro-array (13 sections of original tumor, 13 healthy tissues). Cross-cancer activity was evaluated on biopsies from patients with solid tumor or blood cancer: CRC (n=63), NSCLC (n=30), Osteosarcoma (n=4), pancreatic (n= 48), GIST (n=2), AML (n=10), DLBCL (n=7). Xenograft mice model: XON7 efficacy was tested in several sub cutaneous xenograft models in NMRI nude mice (n=10 per group) using A549: non-small-cell lung cancer, HCT-116: colon cancer, LnPac: prostate cancer, KMS-12-BM: myeloma, T1301: T cell lymphoma. Treatments (35mg/kg, twice a week) were given for 4 weeks once the tumor reached 50 mm3. Results: XON7 induced specific human tumor cell lines CDC (IC50=50ug/mL) and apoptosis (IC50= 100ug/mL) up to 100% of the cells, without cross-reactivity with healthy human cells. XON7 targets several patients’ tumoral tissues. In vivo, XON7 induces a significant reduction of tumor growth ranging from 40 to 90% across all tested tumors. Conclusions: Immunotherapy with XON7 could be a novel therapeutic approach to fight solid tumors and hematological malignancies. [Table: see text]
An engineered Trop2/CD3 bispecific antibody for treatment of gastric, pancreatic, and colorectal cancers.
Authors: O Diana , Chow Kronos , Iyer Deepak , Wong Monarca , Wong Tony K , Wong Dennis A , Luk John Moon
Published: June 2023

e14521

Background: Given its over-expression and poor prognosis in a variety of solid tumors, trophoblast cell surface antigen 2 (Trop2) is regarded as a potential therapeutic target. Trop2-directed antibody-drug conjugates have been developed for treatment of solid tumors; however, their clinical benefits can be limited by the associated undesirable effects which is likely attributed to the on-target off-tumor effect arising from the low basal Trop2 expression on the surface of multiple normal epithelial tissues. The present study aims to develop a humanized Trop2/CD3 bispecific T-cell engager antibody that can promote immunological synapse formation with tumor cells overexpressing Trop2, redirecting T cell-mediated killing specifically to cancers with reduced risk of on-target off-tumor effects. Methods: A Trop2/CD3 bispecific antibody was generated by linking a humanized anti-Trop2 monoclonal antibody with a CD3-binding scFv. To tune the affinity of the bispecific antibody for tumor target, in silico paratope prediction was firstly done using ProABC-2 (Ambrosetti F et al., 2020), following by alanine substitution of the identified paratope residues individually. A total of 6 CDR variants were generated with their binding to Trop2 examined using biolayer interferometry. Selected candidates were further characterized for their binding to Trop2-positive cancer cells and activation of T cells upon target engagement. Redirection of T cell-mediated killing on cancer cells expressing high Trop2 level was also studied in vitro. Results: Substitution of a complementarity-determining residue (CDR) on the heavy chain of anti-Trop2 antibody with an alanine significantly reduced binding affinity to Trop2. This CDR variant was selected for further functional characterization. Apart from weaker binding to Trop2-positive tumor cells, the variant also demonstrated a slower cytokine induction, a less acute T-cell activation, and a cytotoxic effect which was more restricted to Trop2-high cancer cells. Conclusions: Our new Trop2/CD3 bispecific antibody demonstrates potent anti-tumor activity, while at the same time shows a reduced risk of on-target off-tumor toxicity. The drug candidate warrants further investigations including toxicity study in monkeys. [Table: see text]
Tolerability and response of palliative radiotherapy in patients receiving tumor-infiltrating lymphocyte therapy.
Authors: Rogers Max , McKenzie Grant , Rattani Abbas , Yusuf Mehran , Chesney Jason Alan , Dunlap Neal E.
Published: June 2023

e14527

Background: Tumor-infiltrating lymphocyte (TIL) therapy is an emerging therapeutic option for patients with metastatic tumors having progressed on multiple lines of systemic therapy. It is unknown how TIL therapy impacts the tolerability and effectiveness of palliative radiotherapy (RT). We herein present a case series of patients having received both TIL therapy and palliative RT along with the associated local responses and toxicity profiles. Methods: A single institution retrospective database review identified patients treated with TIL therapy following progression on ≥2 standard treatment(s) for metastatic disease. Patients were eligible for inclusion if they received palliative radiotherapy treatment within 1 year of TIL delivery as pre- or post-TIL therapy. Patient demographic data was collected and descriptive statistics reported. Radiographic imaging response was assigned in accordance with Response Evaluation Criteria in Solid Tumors (RECIST). Toxicities were graded in accordance with Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0. Results: Between 2017–2021, a total of 10 patients receiving TIL therapy treated across 12 courses of palliative RT were included for analysis. 8 of 10 patients (80%) had a primary diagnosis of malignant melanoma and 2 (20%) had non-small cell lung cancer. RT was administered to 7 patients pre- and 3 patients post-TIL therapy. Within the pre-TIL cohort, 2 patients received RT between TIL harvest and infusion. Patients received an average total dose of 26 Gy (range, 8-45 Gy). Median patient follow-up after RT was 6 months. Of the patients who received RT prior to TIL therapy, 6 of 9 sites demonstrated at-least stable disease at the site of RT versus 1 of 3 sites receiving post-TIL RT. Across the cohort, 2 patients experienced Grade 1 toxicities associated with RT treatment. No Grade 2+ events were reported. Conclusions: Palliative RT appears to be safe and tolerable when delivered both prior to and post-TIL therapy. RT may be an option for bridging therapy between TIL harvest and infusion. Local response rates to RT were greater in patients receiving RT prior to TIL infusion. Further investigations are warranted to determine the optimal dose, fractionation, and timing of RT when integrating it into future TIL therapy protocols. [Table: see text]
Weight loss (WL) as a predictive factor for immune oncology (IO).
Authors: Fugere Brad , Chen Jim Zhongning , Mazahreh Farah , Fugere Tyler , Mazahreh Liyan , Safar A. Mazin
Published: June 2023

e14528

Background: WL is an adverse prognostic factor/symptom reported in ~40% cancer (ca) patients (pts) at diagnosis agnostic to site or histologic grade. The mechanism of WL in ca, however, remains elusive. The role of energetic waste in ca WL is doubtful (it neglects likely dietary compensation). On the other hand, lymphocytic activation, a clonal, energetically demanding process is accompanied with anorexia limiting caloric intake which leads frequently to WL if protracted over months as seen in lymphoma (B-symptom) or indecisive immune engagement (Eng) with tuberculosis. WL of > 5% of body weight, was an adverse prognosticator and exclusion criterion in non-small cell lung cancer (NSCLC) chemotherapy (CT) trials, likely due to CT or steroids' abrogation of the putative, limited immune ca containment. We hypothesized that WL in ca reflects months of immune Eng, ineffective as it may be at containing the tumor, but nevertheless representing immune Eng with the ca. Such cases stand the best chance for IO benefit. Notably, IO impact in ca mirrors the 40% of WL in ca pts. Methods: Tumor registry retrieved data from all stage 4 cancers treated with monotherapy IO in our institution. We dichotomized pts to beneficiaries (B) or non-beneficiaries (NB). Bs survive longer than median survival for the respective stage/line of therapy in each ca derived from published trials. To account for missing WL data prior to IO, we reasoned that weight gain of ≥10 lbs (reliably charted over 1 year from IO infusions) is acceptable surrogate for WL and met our criteria. Results: WL in our cohort (n = 53) of Melanoma, NSCLC, renal cell carcinoma, and bladder ca was 43% of B; 10% in NB (ratio > 4:1) rejecting the null hypothesis (p < 0.05). PDL-1, Tumor Mutation Burden, both inconclusive in identifying durable benefit, were limited in our set precluding further analysis. Conclusions: Our results support, though do not prove, that WL identifies the appropriate IO pts. Improved outcome despite a historically-adverse prognosticator (WL); the fact that IO was the only ca therapy used, both support our interpretation. We hope to stimulate others to examine independent larger datasets to confirm this notion and to allow comprehensive subset analyses. For WL to be utilized properly the relevant clinical context is crucial, accounting for GI obstructing ca (esophageal ca), enzymatic anomalies (pancreatic ca), volume fluctuations (cirrhosis), catabolism (prolonged hospitalization), or dental issues, to name a few. Additionally, factors such as tumor bulk, perhaps discerned from the T of TNM staging, deserve examination since more vigorous immune activation against larger targets could produce more impact on appetite and caloric intake. Finally, available clinical trial datasets, not tumor registries, might provide a more crisp, reliable source as we continue this investigation. WL can, with confirmatory work, be clinically incorporated as a predictive factor for IO benefit.
"Armed" CEA CAR-T with a SIRPγ-CD28 chimeric co-receptor to exhibit the enhanced antitumor activity in preclinical study of colorectal cancer.
Authors: Zhao Yongchun , Zhu Xiuxiu , Shen Junjie , Xu Yanmin , Yang Zhi , Qi Yanan , Chen Jun , Hong Juan , Wang Linling , Qin Lei , Liu Ge , Huang Xia , Hu Jingwen , Zhao Wenxu , Chen Yunfan , Jiang Xiaorui , Jiao Liurong , Chen Nan , Qian Cheng
Published: June 2023

e14530

Background: Nowadays, CAR-T cell therapy still faces limited therapeutic efficacy for solid tumors. SIRPγ is known as a ligand for a well-known immune checkpoint molecule CD47. It is distinguished from other members of its family in that it has a very short intracytoplasmic tail and is incapable of transducing signals on its own. Of note, the role of SIRPγ still largely unknown to date. Preliminary data indicated SIRPγ plays a key role in T-cell transendothelial migration and promotes antigen-specific T-cell proliferation. We thus designed a novel SIRPγ-CD28 chimeric receptor comprising the extracellular part of SIRPγ, the transmembrane and intracellular domains of CD28 and used as the co-receptor for a CEA-targeting CAR. Here, we report on the activity of this "armed" CAR-T with the SIRPγ-CD28 co-receptor in colorectal tumor (CRC) and its anti-tumor efficacy in mice xenograft model. Methods: In vitro luciferase-based cytotoxicity assay were used by coculturing CAR-T and DLD1-CEA CRC cells to assess the activities of CAR-T w/wo the co-receptor. The incubation supernatants were collected for detecting the release of cytokines. In vivo evaluation of the anti-tumor efficacy of CAR-T with the SIRPγ-CD28 co-receptor versus control T cell, and control CAR-T was performed in NOG mice CRC xenograft model. DLD1-CEA cells expressing luciferase reporter were s.c. implanted and T cells were i.v. injected. Anti-tumor efficacy was assessed by in vivo system. Results: CAR-T with the SIRPγ-CD28 co-receptor exhibited specific cytotoxicity only to CEA+ CRC cells, and the efficacies of CAR-T w/wo the co-receptor were comparable (88.6% vs 87.8%, p > 0.05). However, cytokines were significantly higher in the CAR-T group with the co-receptor. The levels of IFNγ, IL2 and TNFα in groups of CAR-T w/wo the co-receptor were 72,793.33 vs 19,013.33 pg/ml (p < 0.05), 1834.67 vs 25.15 pg/ml (p < 0.05) and 79.65 vs 0 pg/ml (p < 0.05), respectively. In mice CRC xenografts model, the tumors were totally eliminated in the CAR-T group with the co-receptor at 21 days and lasted until the end of experiment, while the tumor growth was suppressed in the control CAR-T group at first and out-of-control in the end (Table). Conclusions: These preclinical studies demonstrated the potential of SIRPγ-CD28 co-receptor as a novel T cellular activation signals for CAR-T. The "armed" CAR-T exhibits a fabulous anti-tumor efficacy in CRC and has great potential for applications in other solid tumors. [Table: see text]
Innovative platforms for development of TCR-T cell therapies against novel tumor targets: Triple negative breast cancer.
Authors: Abbott Rachel Judith Mary , Hammerl Dora , Kortleve Dian , van Brakel Mandy , Wijers Rebecca , Roelofs Daphne , Badoux Madelon , Debets Reno
Published: June 2023

e14536

Background: Adoptive T cells therapies have shown great potential over the past decade, particularly with the use of CAR-T cells to treat haematological tumors. However, their effect in the treatment of solid cancers is limited, due to a lack of relevant therapeutic targets and suppressive mechanisms of the tumor microenvironment. To counteract these challenges, we have developed Pan Cancer T (PCT) platforms to yield novel TCR-T cell therapies against untapped (intracellular) tumor targets exclusively expressed in 10 hard-to-treat solid tumors (including breast, melanoma, colorectal, glioma, lung cancer). Furthermore, we are developing engineering strategies aiming to improve the longevity of T cells within the inhibitory solid tumor microenvironment. Methods: The platforms we have developed to enable generation of genetically engineered TCR-T cells to recognize tumor-specific (intracellular) antigens include: 1) PCT Target Discovery (identifying and selecting safe, immunogenic tumor targets by combining advanced in silico and laboratory techniques); 2) PCT TCR Discovery (identifying, selecting, and validating potent TCR-T cells binding with high specificity and avidity); 3) PCT Next Gen Engineering (identifying drivers of immune evasion and implementing TCR-T cell-based gene-engineering approaches to overcome suppression and/or improve durability). Results: Using our PCT Target Discovery platform, we have validated a set of 30 unique tumor targets which are safe, immunogenic and highly shared. Our first showcase target is PCT-1, which is of particular interest in triple negative breast cancer (TNBC), where it is expressed by >85% of patients, with high homogeneity in >80% of all tumor cells. PCT-1 is also expressed in other tumor types, including >90% melanoma and >45% multiple myeloma patients. The PCT TCR Discovery platform has enabled development of a highly specific and active TCR against PCT-1, which outperformed standard of care TNBC treatments (ie, cis-platinum and sacituzumab govitecan) in advanced preclinical efficacy models, including in vitro killing of patient-derived 3D organoids and an in vivo model in NSG mice. Conclusions: The PCT platforms have enabled development of a promising TCR-T against a novel target antigen expressed in TNBC, an aggressive form of breast cancer that affects younger women with limited treatment options. A portfolio of 29 additional untapped targets has also been identified, providing potential to generate TCR-based therapies for treatment against a much larger range of solid tumors. Furthermore, we are developing next generation technologies, aiming to enhance the durability of responses within the hostile tumor microenvironment, which should be broadly applicable to TCR-T therapies.
Development of allogenic nonviral RNA-based CAR-NK therapy targeting CDH17 in relapsed/refractory gastrointestinal cancer.
Authors: Roy Jaydeep , Lin Vivian , Rahmani Mehran , Chow Kronos , Lin James Chieh-Liang , Chang Alarng , Luk John Moon , Chan Anthony Chun Fung
Published: June 2023

e14539

Background: Gastrointestinal (GI) cancers are commonly fatal after metastasis. The incidence and mortality of GI tumors are high, and the benefits of existing therapies are limited. Cadherin-17 (CDH17) is highly prevalent and specific to GI cancers which has potential for the development of novel treatments for GI cancers. Cell-based immunotherapy using genetically engineered autologous Chimeric Antigen Receptors T (CAR-T) cells has been clinically efficacious for treating hematological malignancies. However, its manufacturing is extremely expensive and lengthy with ~10% failure production rate. Equipping NK cells with CAR may represent a safer and equally effective approach for use in solid tumors as an alternative to autologous CAR-T-cell therapy. Nonviral RNA-based expression of CAR vector in NK cells will provide immediate and multiple accessibility to cancer patients, lower risk of abnormal genetic alteration mediated by viral integration, and potentially limit side effects. This paper demonstrates the development of RNA-CAR-NK targeting CDH17-positive GI cancers. Methods: Ex vivo expanded human peripheral blood NK (Magicell-NK, Medigen, Taiwan) cells were used in this study. To generate anti-CDH17 (C7) CAR-NK cells, mRNA containing a novel CDH17-specific single-chain variable fragment and the 4-1BB costimulatory domain was electroporated to Magicell-NK cell. Flow cytometry analysis was performed to evaluate the cell viability and the expression level of CAR and cytotoxic surface markers of the CAR-NK cells. CDH17 specific cytolytic activity against AGS gastric cancer cell line and the concentration of cytokines released by CAR-NK cells were measured by in vitro cytotoxicity assay and cytokine release assay, respectively. Results: Our results demonstrated efficient electroporation with high-level C7 CAR expression ( > 80%) and optimal CAR-NK cell viability (~60%). Interestingly, 12 h co-culture of C7 CAR-NK cells with AGS gastric cancer cells at 1:1 and 2:1 effector/target (E:T) ratio showed a significantly enhanced antitumor response of C7 CAR-NK cells on AGS-CDH17+ cells but not on AGS-CDH17- cells, indicating CDH17 target-specific cytolytic activity. Moreover, we observed an elevated level of cytokines such as INF-γ and TNF-α released by C7 CAR-NK cells only in co-culture with AGS-CDH17+ cells, confirming the mechanisms behind the CAR-mediated cytotoxicity. Furthermore, these results were supported by an increased level of surface CD107a or LAMP-1 expression in C7 CAR-NK cells during the 12 h of co-culture with AGS-CDH17+ cells at 1:1 and 2:1 E:T ratio. Conclusions: Our findings suggest that a non-viral based CDH17 CAR-NK cell therapy could be a potential allogeneic, off-the-shelf CAR-NK therapy candidate in GI cancer and support the rationale of further investigating it in vivo and clinical trials.
Prognostic impact of baseline circulating extracellular vesicles (BCEVs) in the setting of patients with advanced non-small cell lung cancer (ANSCLC) eligible for first line treatment with osimertinib.
Authors: Gristina Valerio , Barraco Nadia , Taverna Simona , Manno Mauro , Raccosta Samuele , Lamantia Maria , Carreca Anna Paola , Incorvaia Lorena , Badalamenti Giuseppe , Galvano Antonio , Bazan Viviana , Russo Antonio , Carreca Ignazio Ugo
Published: June 2023

e14553

Background: In the liquid biopsy era, the interrogation of extracellular vesicles (EVs) content might be complementary for cancer prognostication and response assessment. In this explorative study, we aimed to describe whether the serial characterization of BCEVs plasma tumor-derived EVs could longitudinally reflect response and resistance to available first-line treatments, investigating the potential to predict clinical outcomes in patients with advanced NSCLC (aNSCLC). Methods: From 2020/02 to 2022/05, 27 treatment-naïve patients with aNSCLC (stageIIIB/IV), eligible for first-line treatment with an EGFRTKI (Osimertinib) or ICIs (e.g. Pembrolizumab ± platinum‐based chemotherapy) based on the predictive molecular pathology, were enrolled. Blood samples (∼5 mL) were collected into K2EDTA tubes early in the morning with fasting condition. We isolated EVs from 2 ml of plasma using exo-Easy Maxi Kit (Qiagen-USA). EVs’s size and distribution was determine by use of Dynamic Light Scattering method (DLS). Besides a Bradford assay (cfEV) for the indirect quantification of EV amount based on EVs proteins content was performed. According to the radiologic response, EV analyses were evaluated in patients with available plasma samples from baseline (T0) to the first radiologic evaluation of disease within 12 weeks (T1). Results: From 2020/02 to 2022/05, 27treatment-naïve patients with aNSCLC (stageIIIB/IV), eligible for first-line treatment with an EGFRTKI (Osimertinib) or ICIs (e.g. Pembrolizumab ± platinum‐based chemotherapy) based on the predictive molecular pathology, were enrolled. Blood samples (∼5 mL) were collected into K2EDTA tubes early in the morning with fasting condition. We isolated EVs from 2 ml of plasma using exo-Easy Maxi Kit (Qiagen-USA). EVs’s size and distribution was determine by use of Dynamic Light Scattering method (DLS). Besides a Bradford assay (cfEV) for the indirect quantification of EV amount based on EVs proteins content was performed. According to the radiologic response, EV analyses were evaluated in patients with available plasma samples from baseline (T0) to the first radiologic evaluation of disease within 12 weeks (T1). Conclusions: This real-world study confirmed the in vivo feasibility of isolating and characterizing BCEVs via DLS and Bradford assay from the plasma of patients with ANSCLC. The quantitative changes of the plasma EVs correlated with survival in the real-time monitoring of such first-line patients, mostly in the oncogene-addicted population receiving osimertinib.
The clinical impact of soluble natural killer cell group 2-member D (NKG2D) receptor ligands on tumor tumorigenicity and anti-tumor immunity.
Authors: Serritella Anthony V. , Saenz-Lopez Pablo , Dhar Payal , Liu Sizhe , Wu Jennifer
Published: June 2023

e14557

Background: NKG2D is an activating receptor expressed on all human natural killer (NK) cells and CD8 T-cells. It is widely accepted that human tumors shed soluble NKG2D ligands, such as soluble major histocompatibility complex-I chain-related molecule (sMIC), to evade immune activation. Harnessing the NKG2D/NKG2D-ligand axis has emerged as a promising avenue for immunotherapy. However, controversy exists over whether soluble NKG2D ligands are immunosuppressive or immunostimulatory. Methods: Cell lines for melanoma (B16F10), Lewis lung carcinoma (LLC1), and prostate cancer (TRAMP-C2, T-C2) were used as parental cell lines in this study. Derivative cell lines over-expressing the soluble human NKG2D ligand sMICB (B16F10-sMICB, TC2-sMICB and LLC-sMICB) were created. We inoculated these 3 syngeneic transplantable tumor models and their respective parental lines into mice and compared tumor incidence/growth over time. We also compared surface markers representative of cell stemness (SCA-1, CD166 and CD133) in mouse cancer cells between the sMICB-expressing tumor cells and their respective parental cell lines. We then characterized the tumor immune microenvironment by flow cytometry. Results: B16F10-sMICB mice demonstrated a significant delay in tumor development compared to control mice (23 vs. 6 days, p < 0.05). Over longer follow-up, rapid tumorigenesis still occurred in the experimental group, with a 50% tumor incidence by day 26. By contrast, the TRAMP-C2-sMICB mice demonstrated higher tumorigenic ability (p < 0.05). LLC-sMICB mice demonstrated no difference in tumorigenicity. Time to tumor onset was more strongly associated with expression of stemness-like markers rather than sMIC. Once tumors were established, they grew significantly more aggressively in the TRAMP-C2-sMICB, LLC-sMICB and B16F10-sMICB lines vs. their parental counterparts. sMICB expressing tumors contained significantly lower percentages of NK cells and CD8 T-cells in tumor infiltrates than their respective parental lines. NK cells and CD8 T Cells from sMICB-expressing tumors had an impaired capacity to produce interferon gamma. Conclusions: Expression of a soluble NKG2D ligand had minimal impact on tumor establishment but facilitated more aggressive growth of established tumors due to immune suppression. Soluble NKG2D ligands imprint negative anti-tumor immunity onto established tumors. Our data differentiated the impact of NKG2D ligands on tumor onset vs. tumor progression and reinforced the translational concept that soluble NKG2D ligands are strong therapeutic targets for future study.
Implications of GITR expression profile for precision immunotherapeutics.
Authors: Moussa Peter , Kurzrock Razelle , Nishizaki Daisuke , Miyashita Hirotaka , Lee Suzanna , Nikanjam Mina , Eskander Ramez Nassef , Patel Hitendra , Botta Gregory P. , Pabla Sarabjot , Nesline Mary K , Conroy Jeffrey M. , DePietro Paul , Sicklick Jason K. , Kato Shumei
Published: June 2023

e14561

Background: Glucocorticoid-induced TNFR-related protein (GITR) is a type I transmembrane protein expressed mostly on CD25+CD4+ T-regs and upregulated on all T-cells upon activation. It serves as a T-cell co-stimulatory receptor and its role in cancer treatment is being explored. While pre-clinical data using combination therapies involving GITR agonism showed promise, early clinical trials have not shown improved outcomes to date. Methods: RNA expression levels of 397 genes in 514 patients with a variety of cancer types were assessed and analyzed at OmniSeq. Transcript levels were normalized to internal housekeeping gene profiles and ranked as percentiles using a reference population. High expression was defined as ≥ 75th percentile, and low-moderate as <75th percentile. Univariate p-values were calculated using Welch t-test or Fischer’s exact test. Multivariate p-values were calculated using logistical regression. Results: A total of 99 patients (19.3%) had GITR high expression. Cancers with the highest proportion of GITR high expression were lung (7/20, 35%) and breast (19/49, 39%), and both these cancers showed a statistically significant association with GITR high expression on multivariate analysis (lung: OR 3.5, p = 0.04, 95% confidence interval (CI) 1.06 – 10.06 CI and breast: OR 4.59, p <0.001, 95% CI: 2.11 – 9.85). 30% of lung and 30.6% of breast cancer patients had a GITR high with ligand low-moderate expression profile; these cancers had the highest proportion of patients with this profile. GITR high expression showed significant association with high RNA expression of multiple known immune checkpoint proteins (PD-1, PD-L1, CTLA4, LAG3, OX40, ICOS and CD137, but not TIM3) as well as PD-L1 IHC ≥1 on univariate analysis, but statistically significant association was only seen with PD-L1 IHC ≥1 (p = 0.007) and OX40 high expression (p = 0.001) on multivariate analysis. Conclusions: While clinical trials with GITR agonism are ongoing, our data suggests selection of certain types of malignancies depending on GITR status may be required. Moreover, there is a correlation between high GITR and high OX40 RNA expression, which could imply increased efficacy using this targeted combination. Further clinical development based on this immune-profiling is warranted. [Table: see text]
PD-L1 in head and neck cancers: Audit for tertiary care centre in India.
Authors: Peelay Zoya Ravish , Mittal Neha , Patil Vijay Maruti , Singh Ajaykumar Chandrabhan , Noronha Vanita , Prabhash Kumar , Menon Nandini Sharrel , Shah Minit Jalan
Published: June 2023

e14565

Background: There is no data on the expression of Programmed Death Ligand 1 (PD-L1) in Head and Neck cancers (HNC) from the Indian population. Hence, this audit was done to estimate the incidence of PD-L1 expression in HNC’s and to detect factors affecting the same. Methods: Data was collected from 466 cases who were advised for PD-L1 gene expression testing from the HNC OPD of Tata Memorial Hospital, Mumbai. Ethics committee approval for this retrospective audit was obtained. Tumor Proportion Score (TPS) scoring for PD-L1 expression was performed using the VENTANA SP263 PD-L1 Assay. PD-L1 status was determined by the percentage of tumor cells with any membrane staining above the background. The demographic details and factors such as Eastern Cooperative Oncology Group (ECOG) performance status (PS), tobacco use, site of tumor, histopathology and time of biopsy. Descriptive statistics were used for analysis. Factors affecting PD-L1 gene expression were sorted using logistic regression analysis. Results: 466 patients were evaluated with a median age of 49 years (Range:16-85), PD-L1 expression was divided as scores and was observed that TPS-0% (Tumor proportion score) was seen in 73 patients (15.7%), TPS >0-20% was seen in 174 patients (37.3%), TPS >20-50% was seen in 88 patients (18.9%) and TPS >50% was seen in 131 patients (28.1%). Higher expression of PDL-1 was seen in oral cavity as site of tumor (p=0.000), squamous cell histology (p=0.000) and consumption of tobacco (p=0.001). Conclusions: This is one of the first studies evaluating real world data on the expression of PDL-1 in HNC’s in the Indian population and factors affecting it. PDL1 expression of 1% or more is seen in 84.3% of HN cancers in India. The histology of the tumor is the major impacting factor. The data provides novel insight into many factors affecting the expression of PDL-1 in HNC’s and in the future can be of help in developing treatment plans with various immunotherapies. [Table: see text]