bluuebunny/crossref_metadata_2025 huggingface dataset:
Genomic analysis in NF2 meningiomas: Does co-mutation suggest new subgroups?
Authors: Xi Yanfeng , Guo Didi , Han Tiantian , Kong Rongrong , Chen Siqi , Ding Ran , Deng Wanglong , Bu Fanfeng , Cao Shangzhi
Published: June 2023

e14058

Background: Meningiomas are the most common primary central nervous system tumours in the adults, accounting for approximately 30% of intracranial tumours. NF2 alterations are the most common genetic abnormality in meningiomas and are known to initiate events for aggressive-type meningiomas. The purpose of this study was to explore genomic characteristics of co-mutated genes in NF2 meningiomas. Methods: This study retrospectively analyzed the genomic alteration of 347 Chinese Meningioma patients during 2019-2022. Next-generation sequencing (NGS) was performed to detect gene mutations in tumor samples of the patients. Results: NF2 mutations were observed in 160 of 347 patients (46%). Our results showed the NF2-mutant patients tended to have older age compared with NF2-wildtype patients (average age: 56 vs. 47, p < 0.05), and meningiomas were more common in women in both groups. TERT promoter mutation or CDKN2A/B deletion occurred in 16/160 (10%) of all NF2-mutant meningiomas. The proportion of NF2 alterations in tumors that harbored CDKN2A/B deletion or TERT promoter mutation was similar to those without mutations (41.0% vs. 46.8%, χ2＝0.46, p > 0.05). 11 patients defined as NF2-mutant meningiomas were found to co-occurred with POLR2A (n = 3), TRAF7 (n = 3), PIK3CA (n = 2), PIK3R1 (n = 2), AKT1 (n = 1), SMO (n = 1), which were previously reported in a mutually exclusive pattern, identifying unique clinical cohorts. Surprisingly, we observed co-mutation in [ NF2 - TRAF7- PIK3CA] genes in a 43-year-old woman meningioma patient. Conclusions: In summary, NF2-mutant patients were observed in 46% Chinese meningioma population and tended to have older age. Interestingly, POLR2A, TRAF7, PIK3CA, PIK3R1, AKT1 and SMO mutations are reported as exclusive of NF2 mutations, were observed in 11 NF2-mutant patients in this study. The co-mutation of different driver genes in meningioma should be given enough attention, and their clinical significance remain further study.
Mutational landscape of intracranial germ cell tumors.
Authors: Wang Jiangong , Han Tiantian , Wang Mengjuan , Guo Didi , Kong Rongrong , Dong Yuwei , Ding Ran , Chen Siqi , Deng Wanglong , Bu Fanfeng
Published: June 2023

e14060

Background: Intracranial germ cell tumours (GCTs) are relatively rare tumours of the central nervous system, which account for 1% to 3% of cases. The pathogenesis of intracranial GCTs is unknown, molecular diagnostic methods currently play only a minor role in the diagnosis of CNS germ cell tumours, which generally rest on histopathological and secretory features. The objective of this study was to explore the molecular profile associated with Chinese intracranial GCT patients. Methods: This study retrospectively analyzed the genomic features of 24 intracranial GCT patients. Next-generation sequencing (NGS) of 551-gene panel was performed to detect gene mutations in tumor and blood samples of the patients. Results: The tumors originated most frequently in the pineal region (41.6%, 10/24), followed by the sellar tumor (16.6%, 4/24). The average age of the patients was 14 years (range, 4-55years), There is a male preponderance of 2:1. The most common mutation genes were KIT (4.23%, n=6), KRAS (2.82%, n=4) and PTEN (2.11%, n=3). KIT mutations are mainly missense mutations concentrated in exon 17, exon 11 and exon 13. Exon 2 and exon 3 of KRAS gene were the main mutation hotspots, mainly G12X(2.08%) missense mutation. The co-mutation of KIT and KRAS was only in one patient. Unexpectedly, four patients (16.6%) carried germline deleterious mutations, in FANCA(n=1), FANCD2(n=1), FANCE(n=1), SDHA(n=1). Two of the patients with germline mutations co-mutated with KIT and KRAS, respectively. Conclusions: 16.6% of the intracranial GCT patients carry germline deleterious mutations, and 12.5% in Fanconi anemia (FA) gene. Overall, these findings indicate that the Fanconi anemia (FA) pathway and the PI3K/AKT pathway play a significant role in intracranial GCTs tumorigenesis.
Human epidermal growth factor receptor 2/3 (HER2/HER3) expression in higher-grade meningioma.
Authors: Mair Maximilian , Tomasich Erwin , Paiato Christina , Widhalm Georg , Eckert Franziska , Sahm Felix , Hainfellner Johannes A. , Berghoff Anna Sophie , Preusser Matthias
Published: June 2023

e14063

Background: Meningiomas are the most frequently diagnosed primary central nervous system (CNS) tumors in adults. Although most meningiomas can be cured by resection, higher-grade lesions (CNS WHO grades 2 & 3) tend to recur, with limited systemic treatment options after exhaustion of local therapies. Antibody drug conjugates (ADCs) targeting human epidermal growth factor receptors 2 and 3 HER2/HER3) have shown remarkable clinical activity in solid tumors including breast cancer brain metastases and are under investigation in various malignancies. Methods: 64 patients with WHO grade 2 (atypical) and 3 (anaplastic) meningioma have been included. DNA methylation analysis has been performed using Illumina EPIC methylation arrays. Panel sequencing for NF2, PIK3CA, SMO, ARID1A, ARID1B, KLF4, SUFU, TERT promotor, and TRAF7 has been executed applying the Illumina NextSeq 500 platform. HER2 and HER3 expression was evaluated by immunohistochemical analysis using PATHWAY HER2 (4B5, Roche) and HER3 #12708 (Cell Signaling) antibodies on a Ventana Benchmark Ultra staining system. Results: In total, 44/64 (68.8%) patients with WHO grade 2 and 20/64 (31.2%) patients with WHO grade 3 meningioma were included. Median age at surgery was 60 years (range: 19-83), and 37/64 (57.8%) patients were female. DNA methylation classes according to the Molecular Neuropathology classifier were benign-1 in 10 (15.6%), benign-2 in 10 (15.6%), benign-3 in 4 (6.3%), intermediate-A in 26 (40.6%), intermediate-B in 4 (6.3%), and malignant in 10 (15.6%) out of 64 tumors. Weak membranous HER2 expression was observed in 1/64 (1.6%) tumor, which was categorized as WHO grade 3/intermediate-B meningioma characterized by infiltration of adjacent bone and fat tissue as well as positive Ki-67 staining of up to 39% of cells. Similarly, only one sample (1.6%) was HER3-positive which was diagnosed as WHO grade 2 tumor of methylation class benign-2, showing meningothelial morphology with elevated mitotic activity and a mutation in TRAF7. The single HER2+ as well as the HER3+ sample were from two distinct female patients. Conclusions: In our cohort, HER2 and HER3 were only infrequently expressed in a small subset of samples of different WHO grade, methylation class, and genetic background. Exploration of further targets of ADCs which are approved or under investigation in other tumors is currently ongoing.
Medullomyoblastoma: A study of demographic and socioeconomic factors.
Authors: Patel Sagar , Hsia Beau , Keirns Darby , Silberstein Peter T. , Wu Xinxin
Published: June 2023

e14064

Background: Medullomyoblastoma is a rare cerebellar cancer theorized to be of teratomatous origin and a more aggressive variant of medulloblastoma primarily affecting children under 10 Despite treatment with excision surgery, the survival period is less than 1 year. There is no standard treatment and previous studies have only looked at median follow-up and radiotherapy. A study using National Cancer Database data would be beneficial to examine the impact of demographic factors on risk and treatment. Methods: A retrospective cohort study using data from the National Cancer Database from 2005 to 2019 analyzed 19 patients diagnosed with medullomyoblastoma. The study examined demographic factors such as age, sex, race, Hispanic origin, insurance coverage, distance from facility, and Charlson-Deyo score through descriptive statistics and evaluated trends in incidence through regression analysis. Results: A total of 19 patients were identified in the database with a confirmed diagnosis of medullomyoblastoma between 2005 – 2019, with a slightly declining incidence of patients diagnosed per year (R2 = 0.155). The average age of diagnosis was 13.5 years (SD = 12.5, range = 1 – 44 years), with 52.6% of affected individuals being children under the age of 10. Males and females were affected relatively equally (52.6% and 47.4%, respectively). The top primary site was the cerebellum (84.2%). Most individuals (94.7%) had Charlson-Deyo comorbidity scores of 0. The majority of individuals (68.4%) received treatment as some form of tumor excision surgery (local tumor excision with or without combination of photodynamic therapy, electrocautery, cryosurgery, wide excision, or radical excision). The majority of the patients were non-Hispanic and White (94.7%). A higher percentage of patients were privately insured (68.4%) than those who were uninsured or insured by Medicaid. The majority (84.2%) of patients lived in a metropolitan area. Patients lived an average of 48.6 miles (SD = 57.1, range = 0.6 – 189.5) from the treatment facility. No patients received palliative care. Conclusions: To the best of our knowledge, this is the first NCDB study on medullomyoblastoma, filling a crucial gap in knowledge. Results show the majority of patients are non-Hispanic White children, with primary site at the cerebellum, similar to previous case reports. For the first time, socioeconomic characteristics of medullomyoblastoma patients have been documented, such as the majority of patients being privately insured living in metropolitan areas. Treatment-wise, patients receive tumor excision surgery in the form of localized excisions or combination therapies. Currently, more attention is paid to pediatric cases, so a study on adult incidences would provide a more holistic assessment of the cancer. Further investigation is needed to understand the effect of demographic/socioeconomic factors on rarity, earlier diagnosis, and treatment.
Phase I safety and preliminary efficacy of PM1003, a bispecific antibody targeting PD-L1 and 4-1BB, in patients with advanced solid tumors.
Authors: Xue Junli , Sun Yuping , Li DaPeng , Guo Ye
Published: June 2023

e14511

Background: PM1003 is a VHH-based anti-PD-L1/4-1BB bispecific antibody. The anti-4-1BB VHH arm binds to a unique epitope at the CRD4 region of 4-1BB, activating 4-1BB signals through cross-linking in the presence of PD-L1 that is often upregulated in the tumor microenvironment, thus avoiding harm to healthy tissues. This is the First In Human study of PM1003. Methods: This study was conducted with a standard 3+3 dose escalation design (Part A: single doses of 0.02, 0.1, 1, 3, 6, and 10 mg/kg for DLT evaluation after 3 weeks followed by Q2W for further administration) and dose expansion (Part B). For Part A, the primary endpoint was safety and tolerability with secondary endpoint of ORR per RECIST 1.1. For Part B, ORR per RECIST 1.1 was the primary endpoint and the secondary endpoints were safety and PK. Results: As of February 9, 2023, a total of 17 subjects have received at least 1 dose of PM1003, including 16 subjects in Part A (3 subjects in each of the 0.02, 0.1, 1, 3 mg/kg dose levels respectively and 4 subjects in 6 mg/kg), and 1 subject in Part B (1 mg/kg Q2W). In Part A, no DLTs were observed up to 6 mg/kg. Of the 17 patients, TRAEs occurred in 15 subjects (88.2%), ≥ Grade 3 TRAEs occurred in 5 subjects (29.4%). No patient discontinued PM1003 administration due to TRAE. The most common TRAEs were anaemia (35.3%), neutrophil count decreased (29.4%), white blood cell count decreased (29.4%), platelet count decreased (29.4%), fatigue (23.5%). 11 subjects completed at least one efficacy evaluation in part A. The median duration of PM1003 exposure was 9.9 weeks (range, 3.1-31.1 weeks). The ORR per RECIST 1.1 was 9.1% with a DCR of 54.5%. Preliminary evidence of antitumor activity has been observed in 1 subject with gastric cancer (1 mg/kg Q2W) who was enrolled after previous surgery, chemotherapy, anti-PD-1 and anti-HER2-ADC treatment, and had a partial response of 87.8% in target lesion reduction during the first efficacy evaluation (7 weeks) after PM1003 treatment. T1/2 of PM1003 of 1 mg/kg and 3 mg/kg dose level is 6.3 and 12.8 days, respectively. Receptor occupancy of PD-L1 in the 1 mg/kg and 3 mg/kg cohorts reached saturation at 336 hours after the first single dose and maintained at an almost stable state. Conclusions: PM1003 was well-tolerated up to 6 mg/kg and has shown preliminary anti-tumor activity in a patient who was previously treated with multiple drug modalities including anti-PD-1. The dose-escalation at 10 mg/kg and dose expansion are ongoing. Clinical trial information: ChiCTR2100052887 .
A first-in-human study of MIL95, an anti-CD47 monoclonal antibody (mAb), in patients with advanced solid tumors and lymphomas.
Authors: Zhang Qingyuan , Liu Aiguo , Shi Jianhua , Cai Qingqing , Qu Zerui , Liu Sijun , Song Yuqin
Published: June 2023

e14513

Background: CD47 provides a potent “do not eat me” signal that allows the tumor to escape removal by the innate immune system. MIL95 is a humanized, anti-CD47 lgG4 monoclonal antibody and that mediated blockade of CD47 can induce macrophage phagocytosis of cancer cells. To evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics and antitumor activity of MIL95, we conducted a first-in-human phase 1 trial in pts with advanced solid tumors and lymphomas (Protocol number MIL95-CT101, Beijing Mabworks Biotech Co. Ltd.) at 5 clinical study sites in China. Currently the study is still ongoing. Methods: We enrolled pts aged 18 years or older with advanced solid tumors and lymphomas whose disease had progressed after standard systemic treatments. Pts were required to ECOG PS score of 0-1 and adequate organ functions. Pts were enrolled in this two-part dose-escalation study: Part A, to determine a priming dose at 0.1, 0.3, 0.8, 1 mg/kg; Part B, to determine optimal weekly maintenance dose at 3, 10, 20, 30, 45 mg/kg. Dose escalation was performed using an accelerated titration followed by traditional 3+3 design during a 14-day (Part A) or 28-day (Part B) dose-limiting toxicity (DLT) observation period. Results: As of 18 Dec 2022, 24 pts (median age 54.5 years; 37.5% male; 54.2% ECOG PS 1) were treated: 8 in part A, 16 in part B. The common tumor types were lymphomas (25.0%) and breast (25.0%), lung (12.5%) cancers. MIL95 toxicity was manageable, no DLT was observed, and dose escalation was ongoing at 45mg/kg. The most common treatment-related adverse events (TRAEs) occurring in ≥10% of pts were anemia (37.5%), blood bilirubin increased (37.5%), leukopenia (20.8%), pyrexia (16.7%), and fatigue (16.7%). Most toxicities were Grade 1 or 2. Five pts had ≥Grade 3 TRAEs (Grade 3 anemia in 2 pts, Grade 3 decreased appetite in 1 pt, Grade 3 enteritis infectious in 1 pt, Grade 4 platelet count decreased in 1 pt). 4 pts had SD as a best response, with 1 pt (Cholangiocarcinoma at dose 20 mg/kg) on study for >5 months. Conclusions: MIL95 was well tolerated at 1 mg/kg priming dose following by maintenance doses of up to 45 mg/kg weekly. Further antitumor activity may need to be combined with other therapies. Clinical trial information: NCT04651348 .
Therapeutic effect of a VSIG4-specific mAb EU103 in a preclinical model of patient-derived ascites of ovarian cancer.
Authors: Kim Hye Jeong , Kang Kyung-Jung , Shin Jeamin , Lee Joong Won , Kim Byoung-Gie , Park Sang-Yoon , Jeong Joseph H. , Kwon Byoung S. S
Published: June 2023

e14516

Background: V-set and immunoglobulin domain-containing 4 (VSIG4) is one of the B7 family-related proteins that includes PD-L1, VISTA, and CTLA4 ligand. VSIG4 overexpression is correlated with poor prognosis in patients with ovarian cancer, lung cancer, gastric cancer, high-grade glioma, and multiple myeloma. VSIG4 is also known to function as a receptor for complements and the negative regulation of T cell proliferation. Although VSIG4 is highly expressed in tissue-resident macrophages and tumor-associated macrophages (TAM), its role in the tumor microenvironment (TME) is not fully elucidated. Methods: We previously developed a therapeutic antibody, EU103, in a humanized form of a VSIG4-specific antibody. Most importantly, we demonstrated that EU103 directly acts on the tumor-associated macrophages (TAM) and mechanistically induces repolarization of TAM to tumor killing M1 macrophages and blocks VSIG4-mediated T cell suppression, eventually leading to CD8+ T cell proliferation and tumor suppression in a PBMC-humanized human lung cancer mouse model. Results: Expanding our previous findings with the therapeutic potential of EU103 in a non-small cell lung carcinoma (NSCLC), here, we also proved its therapeutic effect in human ovarian cancer. Firstly, TAMs isolated from ovarian cancer patients express high levels of VSIG4 with immunosuppressive M2 macrophage phenotype. Secondly, EU103 treatment of the TAMs in in vitro studies induces M2-to-M1 conversion and activation of T cells in the ascites leading to ovarian tumor cell killing in a dose-dependent manner. Thirdly, in vivo therapeutic efficacy of EU103 in ovarian cancer, especially through M2-to-M1 conversion, is also verified in both CD34+ cell and peripheral blood mononuclear cell (PBMC) humanized mouse models. Finally, we validate the results above by treating ovarian cancer patient-derived ascites with EU103. Conclusions: Almost 300,000 women are diagnosed with ovarian cancer worldwide each year, and ovarian cancer ranks fifth in cancer deaths among women. In the United States alone, we expect approximately 20,000 new diagnoses and more than 12,000 deaths for this year. Given our findings in this study and clinical significance, we propose here the therapeutic potential of EU103, previously developed by our proprietary antibody discovery and engineering technology, in ovarian cancer.
Hypophosphatemia and thrombocytopenia: A retrospective chart review to identify markers for prediction of CRS and ICANS in CAR T-cell therapy.
Authors: Barker Kenneth , Bertolo Aaron , Uhland Cole , Koza Shannon , Katsanis Emmanuel , Husnain Muhammad
Published: June 2023

e14524

Background: CAR T-cell therapy has transformed the treatment landscape of B cell malignancies. Adverse effects of CAR T include cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). We aimed to identify biomarkers predictive of CRS and ICANS post CAR T-cell therapy. Methods: A total of 34 patients (19 male and 15 female) with a median age of 68 years and ranging from 38 to 80 years old treated with CAR T-cells for B-cell malignancies at Banner University Medical Center/University of Arizona Cancer Center were evaluated. Phosphate (Ph), IL-6, ferritin, lactate dehydrogenase (LDH), and platelet count from each patient were recorded through retrospective chart review. A simple two-sample t test and ANOVA T-test were run to determine statistical significances by comparing the values of each parameter of interest between groups. Results: Of the 34 patients, 19 (56%) developed CRS without ICANS, 8 (24%) had both CRS and ICANS, and 7 (20%) developed neither. Of the 19 patients with CRS without ICANS, 13 (68%) had clinical hypophosphatemia compared to 0% of patients without CRS or CRS+ICANS. Average phosphate (Ph) nadir levels in the patients with CRS were significantly lower at 2.08 mg/dL compared to those in patients without CRS at 3.20 mg/dL ( P= .00007). Patients with CRS + ICANS had even lower Ph levels at 1.99 mg/dL when compared to patients without ( P= .004). Thrombocytopenia at time of infusion was significantly associated with the development of CRS with those patients having an average platelet count of 114 x 109/L compared to 184 x 109/L in patients without CRS or CRS+ICANS (P = .037). Interleukin-6 (IL-6) levels were elevated in all of patients who developed CRS with an average peak level of 102 pg/mL, however, an inadequate number of patients without CRS or CRS+ICANS were sampled thus precluding a statistical comparison in peak IL-6 levels between groups. Conclusions: Hypophosphatemia is associated with the development of both CRS alone and CRS+ICANS after CAR T cell therapy. A proposed mechanism is thought to be related to increased phosphate consumption by CAR T cell induced metabolic activity. Thrombocytopenia at the time of infusion was significantly associated with the development of CRS. Hypophosphatemia and thrombocytopenia show promise as markers for the prediction of CRS and ICANS in CAR T-Cell therapy.
Investigator-initiated phase 1 clinical trial of synthetic TIL: Autologous peripheral T cells engineered with polyclonal TIL-derived TCRs.
Authors: Cheng Zhongping , Wu Wei , Xuan Jiekun , Zhao Yue , Cai Jun , Porter Ely , Chen Xi
Published: June 2023

e14525

Background: The efficacy of tumor-infiltrating lymphocyte (TIL) therapy is limited by the rarity of anti-tumor TCRs and exhausted status of the product. Its improvement should focus on a high frequency of diverse anti-tumor TCRs with stem-like phenotype, which can be achieved synthetically by identifying multiple anti-tumor TCRs from the patient's intra-tumor TCR repertoire and subsequently engineering them into the patient’s peripheral T cells. Such a ‘synthetic TIL’ is cost-prohibitive using existing technologies. To enable a synthetic TIL product, we developed ultra-high throughput TCR gene synthesis and screening methods to synthesize and screen > 50,000 TCRs from scRNA-Seq data from > 30 patients and identified > 100 tumor/NeoAg-reactive TCRs. The gene expression patterns of the T cells carrying such experimentally validated tumor/NeoAg-reactive TCRs gave rise to an algorithm called Interpreter to predict potentially anti-tumor TCRs from scRNA-Seq data. Methods: To translate synthetic TIL to the clinic, we developed a method to generate up to 100 sequence-verified TCR-encoding plasmids (selected by Interpreter) from fresh tumor biopsy in just 3 weeks. Our GMP-grade production uses a collective knock-in approach, where a mixture of up to 100 plasmids (each encoding a distinct TCR) at equal proportion are packaged into AAV which serve as template to precisely insert the TCR genes into a primarily monoallelic locus and simultaneously inhibit the endogenous TCR. This gene editing is characterized by a modified scRNA-Seq method called EntrySeq. In first-in-human, N-of-1 trials, 109 to 6x109 synthetic TIL cells were administered to solid tumor patients with or without lymphodepletion and IL2. An investigator-initiated Phase 1 trial was then opened to evaluate safety, preliminary efficacy, and cellular activity of synthetic TIL and to determine RP2D and MTD. Subjects with relapsed, unresectable locally advanced solid tumors who received at least 1 standard systemic therapy are allowed to enroll in this study. Those patients with no standard treatment regimens available, or those who are unable to obtain effective treatment due to economic conditions are also permitted. Results: EntrySeq confirmed that > 80% of the T cells express only 1 TCR, avoiding mispairing of different TCRs. The frequencies of different TCRs were largely uniform and were stable through plasmid, AAV and T-cell mixtures. Up to 1.5x1010 synthetic TIL cells were produced from a patient’s leukopak. 40 to 60% of the T cells in the product for infusion express an exogenous TCR. In first-in-human trials, the administered T cells showed favorable persistence and safety profile. The Phase 1 trial is ongoing. Conclusions: We established a fast, affordable, and GMP-compliant process to manufacture synthetic TIL products, enabling a Phase 1 trial.
WTX212, an erythrocyte-anti-PD1 antibody conjugate, to demonstrate anti-tumor activities in tumor models and patients with cancer with acquired resistance to immunotherapy.
Authors: Xiaoqian Nie , Liu Yuehua , Chen Yun , Zhang Qi , Tong Xiangmin , Huang Yanjie , Yuan Yang , Zheng Jiawei , Xu Li , Gao Xiaofei
Published: June 2023

e14529

Background: Despite the success of immune-checkpoint blockade (ICB) treatments, a considerable proportion of patients cannot benefit from ICB therapy due to acquired resistance. One major challenge is the systemic suppression of immune cells induced by the tumor. Recent studies have revealed that successful immunotherapy drives new immune responses arising from peripheral immune system instead of reinvigorating the pre-existing responses in the tumor. Here we designed WTX212, autologous engineered red blood cells (RBCs) conjugated with anti-PD1 antibodies capable of activating the peripheral immune system. We also evaluated its anti-cancer activities in anti-PD1 non-responsive tumor models and a first-in-human (FIH) trial. Methods: The anti-tumor efficacy of WTX212 was evaluated in both PD1 responsive (MC38 colorectal cancer) and non-responsive (KP lung cancer and pB3 breast cancer) mouse tumor models (n=7 / group). Mice were monitored twice a week for clinical signs and tumor sizes. The mechanism of action was investigated in splenectomized mice and by analyzing the biodistribution of WTX212 using a radiolabeling method. The immune landscape was also profiled through cytof mass and scRNA-seq analyses in the spleen, tumor and peripheral blood. The FIH trial (NCT05707325) explores safety, pharmacokinetics and mechanism in cancer patients with acquired resistance to ICB. WTX212 was manufactured by covalently engineering anti-PD1 antibody with RBCs isolated from patients. By 6th Feb 2023, 3 patients have been enrolled and received at least one dose of WTX212 (20*1010 cells, IV, Q3W). Results: Treatment of WTX212 significantly suppressed tumor growth up to 100% in anti-PD1 responsive and non-responsive tumor models and activated tumor microenvironment with increased infiltrated T cells. Unlike antibodies, WTX212 primarily distributed to the spleen and vascular system. The anti-tumor effect of WTX212 was abrogated in corresponding splenectomized mice. Compared to anti-PD1 antibody, single-cell analysis identified the immune landscape remodeling in the spleen from WTX212-treated mice, with down-regulation of PD-L1+ myeloid-derived suppressor cells (MDSCs) and up-regulation of effector CD8+ T cells. In the FIH trial, 3 patients have been administered with total eight cycles of WTX212. No treatment-related AEs greater than Grade 2 were observed. WTX212 remained in the peripheral blood day 21 post transfusion. WTX212 rapidly activates peripheral immune systems with decreased MDSCs, Tregs and increased effector T cells in peripheral blood from all patients. Conclusions: WTX212 demonstrated anti-tumor activities by activating the peripheral immune system to overcome resistance to immunotherapy in both preclinical models and human patients. WTX212 is well-tolerated supporting the future clinical trials.
VHH-based chimeric antigen receptor T cells against CD70 to eliminate primary and re-challenge renal cell carcinoma tumors in preclinical study.
Authors: Jiang Pengfei , Xiong Qinghui , Wang Haiying , Deng Tian
Published: June 2023

e14540

Background: CD70 is not expressed in normal tissue, but highly expressed on various hematopoietic and solid tumors, including renal cell carcinoma (RCC). It is an ideal target for cancer therapy and recently was proved by good results in clinical trials of CD70 monoclonal antibody. Methods: We obtained 4 VHH antibody（2A5, 8B4, 11C9 and 9D8）against CD70 by screen nanobody library derived from an antigen-immunized alpaca. Generated a panel of CAR-T cells by transduce retrovirus expressing CAR (chimeric antigen receptor) to human T cells isolated from health donors. These CAR constructs include 4 VHH sequences and a human scFv containing VH and VL derived from cusatuzumab. We evaluated CAR-T cell function in vitro included cell phenotype, proliferation, cytotoxicity, and cytokine release. Furthermore, RCC tumor cell-xenograft NOG mice models were used to test the anti-tumor activity of CAR-T cells in vivo. Results: All anti-CD70 CAR-T cells eliminate CD70-positive tumor cells but not CD70-negative cells in vitro. Although there were no significant differences between the killing efficiency of the CAR-T cells, more abundant IL-2 production in two VHH-based CAR-T cells (11C9 and 2A5) were observed in multi-round co-culture assay. CAR-T cells eradicated RCC tumor cell in our Xenograft model. After infusion, human T cells expanded over 250 folds in 11C9 group and over 30 folds in 2A5 group, but only nearly 6 folds in human scFv CAR-T treated group. In our second in vivo experiment, all mice treated with higher dose CAR-T, the VHH groups maintained tumor free statue much longer time than human scFv group. Cured mice were re-challenged by subcutaneously infuse RCC tumor cells, observed and measured tumor in the next 53 days. For 11C9 CAR-T treated group, 3of 7 mice maintained tumor-free, 1 of 7 mice were tumor-palpable but too small to measurement; For 2A5 treated group only 1 of 7 mice maintained tumor-free. A significantly greater expansion of human T cells in 11C9 group but not 2A5 group after re-challenge. Conclusions: Our results indicate a promising VHH-based CD70 CAR-T cells for RCC therapy, supporting further development for future clinical trial and application.
Serum vitamin B12 levels and response to immunotherapy.
Authors: Turgeman Ilit , Reiner Benaim Anat , Regev Tsur Stav , Chahine Haya , Bar-Sela Gil
Published: June 2023

e14542

Background: Underlying metabolic pathways have a role in regulation of response to cancer immunotherapy. Due to its immunomodulatory properties, we sought to investigate the relationship between elevated serum vitamin B12 (VitB12) and survival among individuals with cancer treated with immune checkpoint inhibitors. Methods: Data from a retrospective chart review were used to identify patients with advanced cancer initiating anti-neoplastic treatment and a concomitant VitB12 measurement (elevated: > 820 ng/L), between 1/2010-1/2022 at our institution. Patients on immunotherapy (IO) and other regimens (control) were compared using Mann-Whitney test for continuous variables, and Chi-square test or Fisher's test for categorical variables. Multivariate Cox regression models were used to assess the effect of VitB12 levels on overall survival (OS) and progression free survival (PFS), with adjustment for potential confounders. Results: A total of 501 patients were included (control, n = 408; IO, n = 93). Groups were well balanced for treatment line (first line, 91.4% vs. 94%), VitB12 levels (elevated, 29.9% vs. 23.7%; mean, 762.4 ng/L vs. 687.6 ng/L) and liver function tests. Cancer diagnoses differed between groups (most common, gastrointestinal vs. lung, p < 0.001). Multivariate analysis revealed that high VitB12 was negatively associated with OS in both groups (control: hazard ratio, HR 1.4, 95% CI 1.01-1.96, p = 0.04; IO: HR 2.74 as sum of linear baseline and interaction effects, in log scale), as were age (HR 1.03, 95% CI 1.02-1.04, p < 0.01), male gender (HR 0.66, 95% CI 0.50-0.88, p < 0.01), and neutrophil to lymphocyte ratio (NLR) (HR 1.05, 95% CI 0.48-0.99, p = 0.01). However, the only negative prognostic factor for PFS was VitB12 in the IO group alone (calculated HR 8.34). No association was observed with patient and cancer characteristics, other laboratory values, chemotherapy (p = 0.3) or biologic therapy (p = 0.8). Mean follow-up in the control and IO groups was 93 vs 54 weeks (OS), and 22 vs 33 weeks (PFS), respectively. Conclusions: Elevated VitB12 levels were negatively associated with response to immune checkpoint inhibitors, independently of other prognostic factors such as age, cancer type, treatment line and NLR. In addition to a direct role in cellular immunity, VitB12 is involved in gut microbiota balance, which in turn affects host immune responses – suggesting a possible immune-metabolic interplay. Further research is needed to elucidate the metabolic regulators of response to immunotherapy and potential strategies of intervention, in order to rationalize and broaden its clinical utility.
Extracellular vesicles budding from stromal macrophages from the blood circulating in patients with metastatic non-small cell lung cancer to predict clinical outcomes.
Authors: Moran Jillian , Adams Daniel L , Edelman Martin Joseph , Lopez Bravo Pablo , Xu Ting , Liao Zhongxing X. , Gardner Kirby P , Tang Cha-Mei , Lin Steven H.
Published: June 2023

e14547

Background: Extracellular vesicles (EVs), which include exosomes, microvesicles, and apoptotic bodies, are involved in cellular communication, aiding tumor growth, and in metastasis of cancer. Recently, extracellular budding structures on Cancer Associated Macrophage-Like Cells (CAMLs), a subtype of phagocytic circulating stromal cells found in circulation, was observed in patients with metastatic non-small cell lung carcinoma (mNSCLC). In this prospective analysis of n=104 metastatic NSCLC (mNSCLC) patients, we enumerated EV budding on CAMLs to determine their clinical significance on Progression Free Survival (PFS) & Overall Survival (OS), further subtyping based on treatment with or without PD-L1 Immunotherapy (IMT) based on standard of care treatment. These preliminary data suggests that EV positive (EV+) CAMLs prognosticates for worse PFS & OS in mNSCLC. Methods: We initiated a single blind multi-year prospective study to investigate the relationship between EV budding in CAMLs to PFS & OS prior to start of new treatment lines for mNSCLC. Anonymized blood was procured from n=104 pathologically confirmed mNSCLC patients and filtered to isolate CAMLs to measure EV budding using tumor/EV markers (i.e. cytokeratin, CD63, or CD81) and an immune specific marker (PD-L1). Blood was filtered by CellSieve microfiltration and EV budding was characterized as small (≤1 µm) bulbous protrusions from the cell cytoplasm. EVs were quantified by presence (EV+) or absence (EV-), as well asPD-L1 positivity or negativity in EVs, to compare PFS & OS with hazard ratios (HRs) at 60 months by censored univariate and multivariate analyses. Results: CAMLs were identified in 93% (n=97/104) of all samples, with EV budding (EV+) identified in 62% (n=60/97) of CAMLs. At 60 month follow-up, it was determined that EV+ CAMLs was associated with significantly worse PFS (HR=1.67 95%CI=0.4-1.0, p=0.0410) and OS (HR=1.88, 95%CI=0.3-0.9, p=0.0108). Further, in patients not treated with IMT, EV+ CAMLs was associated with significantly worse PFS (HR=2.51, 95%CI=0.2-0.8, p=0.0251) and OS (HR=2.32, 95%CI=0.2-0.9, 0.0407). In contrast, in patients treated with IMT, EV+ CAMLs had no correlation to PFS (1.18, 95%CI=0.5-1.6, p=0.7050) or OS (HR=1.53, 95%CI=0.3-1.3, p=0.2858), suggesting a clinical benefit in treating EV+ patients with PD-L1 IMT. Conclusions: EV budding on phagocytic stromal cells found in the blood of mNSCLC patients appears to predict for poorer PFS and OS, which is reduced with additional of PD-L1 immunotherapies. Larger validation studies are ongoing in both mNSCLC and stage III NSCLC patients, along with analysis of PD-L1 EV staining.
Relationship of platelet-to-lymphocyte ratio with tumor immune microenvironment and efficacy of anti-PD-1 combined with neoadjuvant chemotherapy in muscle invasive bladder cancer.
Authors: Hu Hailong , Niu Yuanjie , Li Hong Zheng , An Zesheng , Wu Zhouliang , Shen Chong , Zhang Zhe , Zhao Gangjian , Huang Shiwang
Published: June 2023

e14549

Background: The platelet-to-lymphocyte ratio (PLR) has been used to predict therapeutic response in different tumors. However, no assessments of its usefulness have been performed in muscle-invasive bladder cancer (MIBC) patients receiving anti-PD-1 combined with neoadjuvant chemotherapy. Methods: The respective data of 62 MIBC patients receiving anti-PD-1 combined with neoadjuvant chemotherapy were analyzed. Whether PLR might predict treatment response to anti-PD-1 plus neoadjuvant chemotherapy and the cutoff value of the parameter were determined by ROC curve analysis. Moreover, 31 tumor mRNA sequencing data before the treatment were collected and analyzed using ESTIMATE R package. Results: PLR before the second period (PLR2, cut-off = 122.38, AUC = 0.667, 95% CI: 0.529-0.805, sensitivity = 0.935, specificity = 0.452) seemed to be useful predictors to determine patients who have a response or no response. The PLR2-low and PLR2-high groups were divided according to median score (154.9), and the estimate score and the immune score were significantly higher in the PLR2-low group. Conclusions: The PLR2 allowed for the accurate and efficient stratification of MIBC patients receiving anti-PD-1 plus chemotherapy and is easily applicable for clinical practice at no additional cost. Meanwhile, we identified that the diagnostic performance of PLR2 might reflect the tumor immune microenvironment, which plays an important role in immune therapy.
Dynamic monitoring of ctDNA to predict response to immunotherapy plus chemotherapy in BTC.
Authors: Liu Chen , Xiao Mingzhe , Wu Yingjun , Ao Jianyang
Published: June 2023

e14550

Background: The survival of advanced BTC patients, especially those who received palliative surgery was unfavorable and the treatment options for these patients were limited. Immune checkpoint inhibitors （ICIs） were approved for the treatment for patients with BTC. However, only a subset of patients benefits from this treatment and the clinical performance was not well evaluated. Therefore, there is a need for a method that can promptly and dynamically evaluate the efficacy of patients receiving immunotherapy. Previous studies have shown that ctDNA can detect relapse and monitor therapeutic efficacy in diverse cancer. We performed a retrospective observational study examining whether ctDNA could serve as a biomarker to track treatment responses in BTC. Methods: Patients diagnosed with advanced BTC and who received ICI were screened and enrolled between January 2020 and October 2022 at Shanghai Eastern Hepatobiliary Surgery Hospital. The key inclusion criteria for patients were as follows: a recent diagnosis of gallbladder/intrahepatic cholangiocarcinoma; received immunotherapy (or plus target therapy); received ctDNA NGS monitoring. Patients were excluded based on the following criteria: patients with multiple primary cancers, or had history of allogeneic organ transplantation or allogeneic hematopoietic stem cell transplantation, or HIV infected person, or severe infections with poor clinical control. Blood samples were collected for ctDNA analysis at the beginning, after 3 and 6 months of ICI treatment. Results: The median age was 66 (42-79), and 12 (44.4%) were male. The ECOG score was mostly 1 (23 cases, 85.2%). Most of the patients were stage IV (21, 77.8%), and there were 5 (18.5%) stage III patients, 1 (3.7%) stage II and 0 (0%) stage I patients. The BTC patients were mostly histologically diagnosed with gallbladder cancer (GBC, 11, 40.7%) and intrahepatic cholangiocarcinoma (8 cases, 29.63%). There were 55% patients received palliative surgery. Among all mutations detected in ctDNA, the mutation status of KRSA at baseline was detected to be related to patient survival, and the KRAS-mut group had a significantly lower probability of overall survival compared to the KRAS-WT group (p=0.014). Patients with more than 3 new mutations during treatment presented a significantly unfavorable OS (p=0.025) and a trend of unfavorable PFS (p=0.062). Besides, patients with increasing amounts of total ctDNA during dynamic monitoring showed significantly worse overall survival probability (p=0.023). Conclusions: Mutation of KRAS in tissues at baseline might be a predictive marker of response to immunotherapy and survival prognosis. ctDNA as a compose biomarker can monitor treatment responses and predict overall survival. Considering the small size and the relatively short follow-up time of our cohort, further prospective and randomized controlled studies are indispensable.
Correlation between MSI, TMB, and TGFBR2 gene mutation in solid tumors.
Authors: Fan Xinglong , Hu Huimin , Cao Yufeng , Xuan Tiantian , Xin Jianjun , Gao Qiangfang , Ma Juan , Wang Chenxi , Ding Ran , Bu Fanfeng
Published: June 2023

e14558

Background: TGFBR2 encodes a receptor kinase which signals to downstream effectors in the TGF-ß signaling pathway. Recent research has shown that truncation mutations of TGFBR2 in gastric cancer with high tumor mutational burden. However, the relationship remains unclear in other solid tumors. Methods: We retrospectively analyzed the TGFBR2 mutations of 19370 Chinese patients with pan-cancer during 2019-2022 (Simcere Diagnosis, Nanjing, China) Next-generation sequencing (NGS) assessed somatic mutations in tumor tissue. We figured out TGFBR2 mutation frequency, TMB and MSI in TGFBR2-mutant, TGFBR2 wild-type. Results: We have detected TGFBR2 mutants in 536 (2.8%) samples. The top 5 frequent cancers were colorectal cancer (132, 24.7%), lung cancer (118, 22%), gastric cancer (78, 14.5%), biliary tract tumors(52, 9.7%) liver cancer (33, 6.2%).TGFBR2 variants included missense/indel/insert (238, 44.4%), truncation (219, 40.9%), splicing site variant (23, 4.3%), and CNV (33, 6.2%).The inactive variants include missense, which occurs in the TGFBR2 serine/threonine kinase domain, truncation and splicing site variant. However, truncation was the most common type, recurring in a few hot spots (p.K153Sfs*35, p.K153Afs*3, p.R520* ). And we found that the TMB and MSI in the TGFBR2 mutation group was significantly higher than that in the TGFBR2 wild-type group [TMB-H (TMB ≥10 musts/Mb): TGFBR2-MUT vs TGFBR2-WT, 41.2% vs 9.5%, p < 0.01; MSI-H: TGFBR2-MUT vsTGFBR2-WT, 27.4% vs1.7% p < 0.01]. In the TGFBR2 mutation group, It was also observed that TMB and MSI in the TGFBR2 truncation or splicing site mutant (INACT) group was significantly higher than that in the other mutant types (VUS) group [TMB-H (TMB ≥10 musts/Mb): TGFBR2-INACT vs TGFBR2-VUS, 51.0% vs 27.9%, p < 0.01; MSI-H: TGFBR2-INACT vs TGFBR2-VUS 39.4% vs 10.6% p < 0.01]. And then, we separately analyzed the correlation between MSI, TMB and TGFBR2 gene mutation in the top 5 frequent cancers. Over 60% of patients with TGFBR2 mutations in colorectal cancers have high TMB and MSI values [TMB-H (TMB ≥10 musts/Mb): TGFBR2-MUT vs TGFBR2-WT, 71.3% vs 12.8%, p < 0.01; MSI-H: TGFBR2-MUT vs TGFBR2-WT, 61.2% vs 3.0% p < 0.01]. Conclusions: We analyzed the distribution of TGFBR2 mutants in Chinese patients with solid tumors. Our data showed that TGFBR2 mutant was significantly associated with TMB-H and MSI-H, and this may be a potential molecular marker of immunotherapy. And studies have shown that the combination of TGF-β inhibitors and immune checkpoint inhibitors can increase the killing effect of T cells on tumors. In short, detection of TGFBR2 mutation has certain clinical significance.
Find: Immunomodulation after a fasting mimicking diet analyzed by Nanostring and multispectral flowcytometry in healthy volunteers: A pilot study.
Authors: de Gruil Nadia , Welters Marij JP , Kaart Sam , Pijl Hanno , van der Burg Sjoerd H. , Kroep Judith R.
Published: June 2023

e14560

Background: An increasing number of clinical trials, including our randomized phase III trial (NCT05503108), are investigating the potential benefit of short-term fasting using a fasting mimicking diet (FMD) on the anti-tumor effect of chemotherapy. Preclinical research suggests that FMD promotes infiltration of CD8+ T-cells, while decreasing regulatory T-cells (Treg). Extensive immunoprofiling is needed to investigate the immunomodulatory effects of FMD alone and guide ongoing translational trials on FMD as an adjunct to chemotherapy in patients with cancer. Methods: FIND is a prospective pilot study, which included 9 healthy volunteers who followed a 4-day FMD every 3 weeks, 2 times (NCT04833439). Peripheral blood samples were collected in a fed state (baseline), after overnight fasting (overnight) and directly after the second cycle of FMD (final). Metabolic markers (IGF1, glucose, ketone bodies) to ensure ketosis were measured in the plasma. The RNA from white blood cells was isolated and transcriptomics was performed with the Nanostring IO360 human Pancancer panel, and analyzed by nSolver Advanced analysis, Graphpad Prism and String analysis. Statistical testing was done using Wilcoxon signed-rank tests in R-studio. In addition, 40-plex spectral AURORA flow cytometry for immune cell composition with FlowJo analysis was performed. This study was approved by the local Medical ethical committee (NL76033.058.21). Results: Paired analysis of the Nanostring data identified 56 differentially expressed genes (DEG) comparing baseline and final samples initially, of which 19 genes showed at least 1.2 log2 fold-change in at least half of the participants. String analysis indicated that the DEGs were associated with fatty-acid metabolism, T-cell and myeloid cell function and immune regulatory pathways. Cell typing showed an increase in cytotoxic NK CD56dim cells (p=0.031) and a decrease in Tregs decreased (p=0.063) after FMD. In-depth analysis of gene specific pathway enrichment and immune cell profiling by flow cytometry will be presented. Conclusions: The expression profile of 56 genes in circulating immune cells changed after 2 FMD cycles, next to fatty-acid metabolism affecting the composition and function of both T-cells and myeloid cells and their regulation. Altogether, FMD induced immunological changes that could support cancer treatment and guide immunomonitoring in future trials evaluating the effects of FMD in cancer treatment. Clinical trial information: NCT04833439 .
Prognostic value of systemic inflammatory response in early-stage triple-negative breast cancer.
Authors: Ovcaricek Tanja , Matos Erika , Auprih Masa , Borstnar Simona
Published: June 2023

e14562

Background: Among breast cancer (BC) triple-negative (TN) subtype is the most immunogenic. Host and systemic immune response characterised by tumor-infiltrating lymphocytes (TILs) and immune cells in peripheral blood shape the net anti-tumor immune status. The systemic immune-inflammation index (SII) represents an effective indicator of the peripheral immune status. To date, TILs and SII have been largely examined separately. Therefore, in the present study, we investigated the prognostic value and relationship between pretreatment TILs and SII in a large number of unselected patients (pts) with early (e) stage TNBC. Methods: A total of 301 consecutive eTNBC pts treated with curative intent at Oncology Institute Ljubljana from January 2000 till August 2007 were investigated. Histopathological and clinical data were obtained retrospectively from the medical records. TILs were scored as intensive vs others. SII was determined by platelet × neutrophil/lymphocyte. Univariate and multivariate analyses were performed using the Cox proportional hazard model. The optimal cut-off value of SII was 675 and was evaluated using the receiver operating characteristic (ROC) curve. Results: The median follow-up period was 17,6 years (16.9-18.3), and the entire cohort`s 10-year DFS was 67.5% and 10-year OS 72.1%. The median age was 54 (53-57) years. Most pts were diagnosed node negative (40.7%), with larger tumors (≥ 2 cm, 57.1%), grade 3 (83.5%) and underwent perioperative chemotherapy (cht) (81%). Patients with positive lymph nodes (LN) had higher SII (r=0.21, p=0.003), no correlation between LN and TILs was found. Patients with higher TILs had significantly better OS (HR: 0.40, 95% CI 0.37-0.97, p=00.004), and lower SII predicted significantly better DFS and OS (HR for DFS: 1.65 95% CI 1,03-2.67, p=0.038, HR for OS: 1.72, 95% CI 1.11-2.65, p=0,015). In multivariate analysis, positive LN status retained its independent prognostic value for worse DFS and positive LN and higher SII for worse OS. Conclusions: Our data confirms important role of systemic immune response in the prognosis of eTNBC. The survival of these pts is significantly influenced by the pretreatment SII and TILs, the prognostic value of SII seems to be stronger in our cohort. However, positive lymph node status remains the strongest predictor for worse survival in eTNBC. [Table: see text]