Innate immune activation by checkpoint inhibition in patient-derived lung cancer tissues
Creators: name: Lane, Andrew
name: Fan, Teresa
name: Higashi, Richard
name: Song, Huan
name: Daneshmandi, Saeed
name: Mahan, Angela
name: Purdom, Matthew
name: Pittman, Thomas
name: He, Daheng
name: Wang, Chi
Publisher: name: Zenodo
Year: 2021

description: Although Pembrolizumab-based immunotherapy has significantly improved lung cancer patient survival, many patients show variable efficacy and resistance development. A better understanding of the drug's action is needed to improve patient outcomes. Functional heterogeneity of the tumor microenvironment (TME) is crucial to modulating drug resistance; understanding of individual patients' TME that impacts drug response is hampered by lack of appropriate models. Lung organotypic tissue slice cultures (OTC) with patients' native TME procured from primary and brain-metastasized (BM) non-small cell lung cancer (NSCLC) patients were treated with Pembrolizumab and/or beta-glucan (WGP, an innate immune activator). Metabolic tracing with<sup> 13</sup>C<sub>6</sub>-Glc/<sup>13</sup>C<sub>5,</sub><sup>15</sup>N<sub>2</sub>-Gln, multiplex immunofluorescence (mIF), and digital spatial profiling (DSP) were employed to interrogate metabolic and functional responses to Pembrolizumab and/or WGP. Primary and BM PD-1<sup>+</sup> lung cancer OTC responded to Pembrolizumab and Pembrolizumab + WGP treatments, respectively. Pembrolizumab activated innate immune metabolism and functions in primary OTC, which were accompanied by tissue damage. DSP analysis indicated an overall decrease in immunosuppressive macrophages and T cells but revealed microheterogeneity in immune responses and tissue damage. Two TMEs with altered cancer cell properties showed resistance. Pembrolizumab or WGP alone had negligible effects on BM-lung cancer OTC but Pembrolizumab + WGP blocked central metabolism with increased pro-inflammatory effector release and tissue damage. In depth metabolic analysis and multiplex TME imaging of lung cancer OTC demonstrated overall innate immune activation by Pembrolizumab but heterogeneous responses in the native TME of a patient with primary NSCLC. Metabolic and functional analysis also revealed synergistic action of Pembrolizumab and WGP in OTC of metastatic NSCLC.
description: Data uploads of curated FT-MS data files as .xlsx pivot tables. One file per tissue type. UK2035_OTC_13C6Glc_WGP_Pembro_MS.xlsx : Patient number 2035 (metaststic lung cancer) tissue slices treated with 13C glucose ±WGP, pembrolizumab. Mass spectrometry. UK096_Mf_WGP_6well_2d_pol_13C6Glc_MS.xlsx: Patient number 96 macrophages ±WGP 2D culture. UK096_Mf_3d_pol_13C15NQ_MS.xlsx: : Patient number 96 macrophages ±WGP 3D culture. UK131_OTC_13C6Glc_MS_data.xlsx: Patient number 131 tissue slices 13C Glucose tracer. UK131_OTC_13C15N_MS_data.xlsx : Patient number 131 tissue slices 13C, 15N Glutamine trace. Funding provided by: National Cancer Institute<br>Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000054<br>Award Number: 1P01CA163223-01A1Funding provided by: National Institute of Diabetes and Digestive and Kidney Diseases<br>Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000062<br>Award Number: 1U24DK097215-01A1Funding provided by: National Institute of General Medical Sciences<br>Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000057<br>Award Number: 5P20GM121327Funding provided by: National Cancer Institute<br>Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000054<br>Award Number: P30CA177558
description: The polar extracts were split for NMR and/or IC-UHR-FTMS analysis. 1D <sup>1</sup>H and <sup>1</sup>H(Albers et al.) heteronuclear single quantum coherence (HSQC) NMR analyses were performed on a Bruker Avance III 16.45 T spectrometer or an Agilent 14.1 T DD2 spectrometer outfitted respectively with a 1.7 mm cryoprobe or a 3 mm coldprobe as described previously (K. Sellers et al., 2015). IC-UHR-FTMS analysis was done as described previously (T. W. Fan et al., 2016) on a Dionex ICS-5000 ion chromatography system coupled to a Thermo Orbitrap Fusion Tribrid Fourier transform mass spectrometer with a mass resolving power of 370,000 at 400 m/z. Experimental details of sample preparation are provided in the manuscript (accepted).