Citations

Tumor-Treating Fields (TTFields), a novel therapeutic avenue, is approved for therapy in Glioblastoma multiforme, malignant pleural mesothelioma, and metastatic non-small cell lung cancer (NSCLC). In pancreatic ductal adenocarcinoma (PDAC), several clinical trials are underway to improve outcomes, yet a significant knowledge gap prevails involving the cell-extracellular matrix (ECM) crosstalk. Herein, we hypothesized that treatment with TTFields influence this crosstalk, which is reflected by the dynamic alteration in nanomechanical properties (NMPs) of cells and the ECM in a co-culture system. We employed an ECM gel comprising collagen, fibronectin, and laminin mixed in 100:1:1 stoichiometry to co-culture of Panc1 and AsPC1 individually. This ECM mixture mimics the in vivo tumor microenvironment closely when compared to the individual ECM components studied before. A comprehensive frequency-dependent study revealed the optimal TTFields frequency to be 150 kHz. We also observed that irrespective of the ECM’s presence, TTFields increase cell membrane stiffness and decrease deformation several-folds in both Panc1 and AsPC1 cells at both 48 h and 72 h. Although adhesion for AsPC1 decreased at 48 h, at 72 h it was observed to increase irrespective of ECM’s presence. Moreover, it significantly alters the NMPs of ECM gels when co-cultured with PDAC cell lines. However, AsPC1 cells were observed to be more detrimental to these changes. Lastly, we attribute the stiffness changes in Panc1 cells to the membrane F-actin reorganization in the presence of TTFields. This study paves a path to study complex PDAC TME as well as the effect of various chemotherapeutic agents on such TME with TTFields in the future.

Abstract Background: Pancreatic ductal adenocarcinoma (PDA) is a disease with guarded prognosis with a 5-year overall survival (OS) of 3.1% in patients with metastatic disease. The need for innovative treatments for this lethal disease is paramount. TTFields is a novel treatment modality that employs low intensity, alternating, and intermediate frequency electrical fields that abrogates cancer cell proliferation through different mechanisms. These include inhibition of mitosis, disruption of cancer cell membranes, metabolomic dysregulation of cancer cells, and immunogenic activation of the tumor microenvironment (TME). TTFields are approved in glioblastoma multiforme, non-small cell lung cancer and malignant mesothelioma. The PANOVA trial demonstrated promising efficacy for TTFields plus chemotherapy in patients with PDA. A trial of GCN in patients with PDA demonstrated exceptional efficacy in patients with metastatic disease. Here in, we present our ongoing clinical trial investigating the safety and efficacy of TTFields plus GCN in patients with PDA (NCT04605913). Methods: This a phase I/II trial with an initial safety run-in phase followed by the phase II portion. The primary objective of the trial is to determine the safety (CTCAE v 5.0) of GCN plus TTFields. Secondary objectives include determining the 6-month progression free survival (PFS) rate, overall response rate (ORR, RECIST v1.1), 12-month survival rate, OS, and QOL (EORTC QLQ-C30, version 3.0.2.3). Inclusion criteria include PDA with liver metastasis (patients can have additional sites of metastasis), ECOG 0-1, measurable disease, and ability to operate the Novo TTF-100L (P)/Novo TTF-200T system. Treatment consisted of TTFields (150 kHz 18 hours/day) and GCN (G: 1000 mg/m2, C 30 mg/m2, and N: 150 mg/m2 every 2 weeks, 28 days cycle), followed by a maintenance phase after 6 months (G: 1000 mg/m2 every 2 weeks) plus TTFields until progression. Six patients were to be enrolled in the phase I portion and if ≤1/6 patients experienced a dose limiting toxicity (DLT, treatment emergent adverse event, or skin toxicity of Gr4), the plan would be to move to a phase II trial enrolling 34 more patients. If 2/6 patients experienced a DLT, 6 patients would be enrolled in dose level -1 (GCN plus TTFields for 9 hours/day followed by the maintenance treatment) and if tolerated the phase II portion would start with dose level -1 regimen. The phase I portion was completed without any DLTs and the trial is currently enrolling to the phase II portion. The total number of patients (40) was powered to the 6-months PFS rate and ORR (estimated using the lower bound of exact one-sided 90% confidence intervals). The goal is to "reject" a 6-month PFS rate of 66% and an ORR of 71% (based on previous GCN trial). Time-to-event endpoints will be assessed using the Kaplan-Meier method. Citation Format: Hani M. Babiker, Jason Starr, Jeremy Jones, Conor O’Donnell, Robert Jones, Jacqueline Boni, Lashundra Stewart, Josh Cameron, Oudom Kour, Kabir Mody, Robert R. McWilliams, Daniel H. Ahn, Tanios Bekaii-Saab, Mitesh J. Borad, Umair Majeed. A phase I/II pilot trial, single arm, open label, of gemcitabine, cisplatin, protein-bound paclitaxel (GCN) combined with Tumor Treatment Fields (TTFields) in patient with metastatic pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr CT190.

Sleep is critical to athlete health and sport performance. Purpose To evaluate the relationship between objective sleep data and next-day basketball game performance. Methods Collegiate men basketball athletes (n = 4) wore smart ring sleep trackers the night before games (n = 22). Sleep variables included hours of total sleep (TS), rapid eye movement sleep duration (RM), deep sleep duration (DS), and heart rate variability (HRV). Game performance was evaluated using field goal percentage (FG%) and player efficiency (EFF). Results TS, RM, DS, and HRV demonstrated very weak relationships with FG% ( p > 0.05) and EFF ( p > 0.05). Individual player analysis revealed one athlete exhibited strong to moderate correlations for TS-EFF (r = 0.67), RS-EFF (r = 0.41), TS-FG% (r = 0.51), and RM-FG% (r = 0.67) ( p < 0.05). Sleep metrics had no effect on FG% (R 2 = 0.025) and EFF (R 2 = 0.053) ( p > 0.05). Conclusions Results suggest a lack of group-level associations between smart ring-measured sleep variables and next-day game performance. This highlights the need for individualized sleep monitoring rather than relying solely on objective metrics.

Tumor treating fields (TTFields), a noninvasive anticancer treatment modality, has received FDA approval for pleural mesothelioma, metastatic NSCLC following platinum-based therapy, newly diagnosed and recurrent glioblastoma multiforme. TTFields have also demonstrated potential benefits in other cancer types, including pancreatic ductal adenocarcinoma (PDAC). Although the PANOVA trial yielded positive results in PDAC patients, the mechanisms underlying these beneficial effects remain unclear. Severe desmoplasia is a hallmark characteristic of PDAC due to accumulation of excess extracellular matrix (ECM) caused by activated fibroblasts and cancer cells. This dense ECM contributes to an immunosuppressive tumor microenvironment (TME) and drug resistance, which ultimately leads to poor 5-year survival in PDAC patients. Our previous work with atomic force microscopy (AFM) showed that the nanomechanical properties of the ECM change dynamically based on its composition and the presence of PDAC cells, and vice versa. Herein, we prepared an ECM complex comprising of collagen, fibronectin and laminin at physiological concentrations. Following which, we investigated the alteration of stiffness of both PDAC cells and the ECM in presence of TTFields using an AFM. A frequency-dependent analysis of TTFields showed that a frequency of 150 KHz led to consistent decreases ECM stiffness at 48 and 72 hours. Next, we evaluated the influence of TTFields in PDAC cells alone. In Panc-1 cells exposed to TTFields for 48 and 72 hours, we observed a more than 6-fold increase in membrane stiffness compared to untreated controls. Additionally, AsPC-1 cells subjected to TTFields exhibited a greater than 2-fold increase in membrane stiffness at these time points. Furthermore, we have investigated influence of TTFields on PDAC cells and ECM components in a co-culture model. In this co-culture scenario for Panc-1 cells, TTFields increase membrane stiffness by 5-fold compared to untreated at these time points. Also, AsPC-1 cells demonstrated a 1.5-fold increase in membrane stiffness in the presence of TTFields compared to the untreated. Lastly, we also evaluated the influence of cells on the ECM stiffness in the presence of TTFields. It significantly increased the ECM stiffness in presence of Panc-1 co-culture by 1.3-folds compared to control. We also observed 3- and 4-fold increase in ECM matrix stiffness when co-cultured with AsPC-1 for 48 and 72 hours from TTFields, respectively compared to control. In conclusion, TTFields significantly modulate the stiffness of PDAC cells and the ECM, both individually and in co-culture. Citation Format: Tanmay Kulkarni, Hani Babiker, Debabrata Mukhopadhyay, Santanu Bhattacharya. Tumor treating fields (TTFields) alter nanomechanical attributes of cell-ECM complex in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5105.

Abstract Background: TTFields are non-invasive and novel treatment modality that utilizes low intensity, alternating, and intermediate frequency electrical fields to inhibit cancer cell proliferation through a multitude of mechanisms. These include inhibition of mitosis, disruption of cancer cell membranes, metabolomic dysregulation of cancer cells, and immunogenic activation of the tumor microenvironment (TME). TTFields are approved in glioblastoma multiforme, non-small cell lung cancer and malignant mesothelioma. The PANOVA2 trial demonstrated promising efficacy for TTFields when applied together with chemotherapy in patients with PDAC. Mechanisms of TTFields in PDAC need further delineation. We sought to investigate the efficacy of TTFields in vitro and in vivo in PDAC cell lines and mouse models. Preclinical studies were conducted in concert with an going clinical trial (NCT04605913). Experimental Procedures: Utilizing the inovitro apparatus we studied the effect of TTFields (150kHz, intensity 2.9+0.2 V cm RMS, 37ċ, over 72 hrs) in an array of PDAC (Panc1, KPC, Panc2, and AsPC1) and human pancreatic stellate cells (hPSC) cell lines, alone or together with cisplatin (Cis, 0.1μM) or gemcitabine (Gem, 0.01μM), compared to control. Cell viability analysis was conducted in each of the groups. In addition, cell lysates from Panc1 cells from concomitant treatment of TTFields plus Gem were probed for STAT3 and α-SMA to analyze stromal proteins and RAS pathway components (KRAS, P38, P42, and others) were performed via Westen Blot. We also performed RNA sequencing for the different groups to analyze gene expression levels. We further assessed the effect of TTFields (150kHz, intensity 1.8-2 V/cm RMS, over 2 weeks) using the inovivo system with and without chemotherapy (Cis, 0.25 mg/Kg per dose (i.p.) 3 times a week) compared to control in C57BL6 orthotopic murine models (6-7 weeks old C57BL6 mice). Summary of Results: Our analysis revealed statistically significant abrogation of PDAC cancer cell lines for the different groups and hPSC cell viability when compared to control for TTFields monotherapy (p &lt;0.001) and TTFields concomitantly with Gem or Cis (p &lt;0.0001). Western blot analysis demonstrated that TTFields alone and when combined with chemotherapy alters both the stroma and RAS pathways with confirmed downregulation of pSTAT3 and α-SMA and KRAS, P-P38, and P-P42. In vivo analysis revealed significant decrease in tumor volume and weight for the concomitant effect of TTFields + Cis. Conclusion: Our preclinical studies demonstrated the efficacy of TTFields as monotherapy and concomitantly with chemotherapy in PDAC and hPSC cell lines and mouse models. Mechanistic insights from the pre-clinical studies will inform the design of future TTFields studies in PDAC. Citation Format: Hani M. Babiker, Santanu Bhattacharya, Douaa Albelal, Shamit Kumar Dutta, Enfeng wang, Debabrata Mukhopadhyay. umor treating fields (TTFields) plus chemotherapy abrogates pancreatic ductal adenocarcinoma (PDAC) proliferation and alters the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1834.

Background: Multicellular programs in the tumor microenvironment (TME) drive cancer pathogenesis and response to therapy but remain challenging to identify and profile clinically. Here, we present Spatial EcoTyper, a multimodal machine learning framework for large-scale profiling of spatially dependent cell states and multicellular ecosystems, termed spatial ecotypes (SEs). Methods and Findings: By integrating five million single-cell and spot-level spatial transcriptomes from 10 diverse human neoplasms, including carcinomas and melanomas, we identified a rich tapestry of conserved regional plasticity across nine TME cell types. We then applied Spatial EcoTyper to deeply dissect these data, revealing nine novel SEs with broad conservation - each with unique biology, TME cell states, geospatial features, and clinical outcome associations. SEs were linked to specific biogeographic domains, including the tumor core, margin, and adjacent stroma, and were remarkably well-validated in 235 held-out tumor samples profiled by either single-cell RNA sequencing, sequencing-based spatial transcriptomics (10x Visium), or high-resolution imaging-based spatial transcriptomics (Vizgen MERSCOPE and NanoString CosMx). By deconvolving SEs from over 1,200 bulk tumor RNA-seq profiles, we discovered striking associations between SE levels and response to immune checkpoint inhibitors (ICIs), outperforming nearly 40 potential correlates of ICI response, including previously described signatures of tertiary lymphoid structures and tumor tissue PD-L1 expression. Notably, baseline levels of a proinflammatory SE - localized to the tumor core and defined by a hub of unique T cell, macrophage, fibroblast, and endothelial cell states - strongly forecasted ICI benefit, while another SE - localized to the tumor margin and defined by myofibroblast and hypoxia-associated endothelial cell states - portended ICI resistance. Conclusion: Our data reveal fundamental units of TME organization and demonstrate a platform for large-scale profiling of spatial cellular ecosystems in any tissue, with implications for improved forecasting of immunotherapy response and the development of novel therapeutic strategies. Citation Format: Wubing Zhang,Erin L. Brown,Abul Usmani,Noah Earland,Hyun Soo Jeon,Irfan Alahi,Nicholas P. Semenkovich,Janella C. Schwab,Chloé B. Steen,Chloe M. Sachs,Bilge Gungoren,Faridi Qaium,Peter K. Harris,Jacqueline Mudd,Qingyuan Cai,Antonella Bacchiocchi,Ryan C. Fields,Mario Sznol,Ruth Halaban,David Y. Chen,Aadel A. Chaudhuri,Aaron M. Newman. High-resolution profiling of the tumor microenvironment with spatial ecotypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 153.

ABSTRACT Purpose Pulmonary toxicity (PT) remains a concern following total body irradiation (TBI). This study describes the incidence and factors associated with PT in a cohort of patients homogeneously treated with the lateral opposed fields technique. Methods Medical records of 61 patients ≤21 years of age treated with TBI as a component of stem cell transplantation from 1993–2017 were retrospectively reviewed. The incidences of PT, graft‐versus‐host disease (GVHD), and other toxicities were recorded for each patient. PT was subdivided into its etiologies, and symptomatic PT was established by clinical diagnosis. The association of PT with other variables was examined using chi‐square tests, and overall survival (OS) was estimated using Kaplan–Meier. Results Three‐year OS was 65% (54%–79%). Median TBI dose rate was 7.7 cGy/min (4.0–10 cGy/min). PT occurred in 20 patients (33%). A higher rate of PT was observed in patients with GVHD (65% vs. 35%, p ≤ 0.01). TBI dose rate (</≥ 7.7 cGy/min) was not associated with the risk of PT; however, on continuous analysis, a higher dose rate increased the risk of PT (HR = 1.9, p = 0.04). PT (HR = 3.4, p = 0.01), noninfectious PT (HR = 5.0, p ≤ 0.01), and pneumonitis (HR = 4.3, p = 0.01) were associated with inferior OS. Conclusions This cohort of patients treated with a low dose rate, bilateral technique, and total dose of 1000–1320 cGy experienced an acceptable incidence of PT, comparable to other published studies. Because PT correlated with reduced survival, current practice should seek to address factors associated with PT by aiming to prevent GVHD and by choosing low dose rates, such as in the range of 4.0–10 cGy/min.