Heterogeneous immunogenomic features and distinct escape mechanisms in multifocal hepatocellular carcinoma.

BACKGROUND & AIMS: The presence of multifocal tumors, developed either from intrahepatic metastasis (IM) or multicentric occurrence (MO), is a distinct feature of hepatocellular carcinoma (HCC). Immunogenomic characterization of multifocal HCC is important for understanding immune escape in different lesions and developing immunotherapy. METHODS: We combined whole-exome/transcriptome sequencing, multiplex immunostaining, immunopeptidomes, T cell receptor (TCR) sequencing and bioinformatic analyses of 47 tumors from 15 patients with HCC and multifocal lesions. RESULTS: IM and MO demonstrated distinct clonal architecture, mutational spectrum and genetic susceptibility. The immune microenvironment also displayed spatiotemporal heterogeneity, such as less T cell and more M2 macrophage infiltration in IM and higher expression of inhibitory immune checkpoints in MO. Similar to mutational profiles, shared neoantigens and TCR repertoires among tumors from the same patients were abundant in IM but scarce in MO. Combining neoantigen prediction and immunopeptidomes identified T cell-specific neoepitopes and achieved a high verification rate in vitro. Immunoediting mainly occurred in MO but not IM, due to the relatively low immune infiltration. Loss of heterozygosity of human leukocyte antigen (HLA) alleles, identified in 17% of multifocal HCC, hampered the ability of major histocompatibility complex to present neoantigens, especially in IM. An integrated analysis of Immunoscore, immunoediting, TCR clonality and HLA loss of heterozygosity in each tumor could stratify patients into 2 groups based on whether they have a high or low risk of recurrence (p = 0.038). CONCLUSION: Our study comprehensively characterized the genetic structure, neoepitope landscape, T cell profile and immunoediting status that collectively shape tumor evolution and could be used to optimize personalized immunotherapies for multifocal HCC. LAY SUMMARY: Immunogenomic features of multifocal hepatocellular carcinoma (HCC) are important for understanding immune-escape mechanisms and developing more effective immunotherapy. Herein, comprehensive immunogenomic characterization showed that diverse genomic structures within multifocal HCC would leave footprints on the immune landscape. Only a few tumors were under the control of immunosurveillance, while others evaded the immune system through multiple mechanisms that led to poor prognosis. Our study revealed heterogeneous immunogenomic landscapes and immune-constrained tumor evolution, the understanding of which could be used to optimize personalized immunotherapies for multifocal HCC.

[Pollution Characteristics and Source Apportionment of Atmospheric Volatile Organic Compounds in Summer in Yuncheng City].

Based on the online monitoring data of VOCs, O3, and NO2 in Yuncheng City from June to August 2020, the pollution characteristics of VOCs in Yuncheng City in summer were analyzed. At the same time, the main emission sources were determined using a PMF model, and the chemical reactivity of VOCs was evaluated using the maximum incremental reactivity (MIR) method and fractional aerosol coefficients (FAC). The results showed that the urban area of Yuncheng was seriously polluted by VOCs and NO2 in the early morning and evening during summer, the peak value of VOCs daily variation occurred at 08:00 and 20:00, respectively, and was mainly affected by the morning and evening peaks in traffic. The ρ(VOCs) from June to August was 50.52 µg·m-3, and the species with the highest proportion were alkanes (39.39%) and oxygenated volatile organic compounds (OVOCs, 34.63%). Five VOCs emission sources were determined by the PMF model, of which the largest contribution was from motor vehicle exhaust emission sources (33.10%), followed by industrial emission sources (29.46%), natural gas and coal combustion sources (17.31%), solvent use sources (11.94%), and plant emission sources (8.19%). Controlling motor vehicle exhaust emission sources is the key to alleviate VOCs pollution in summer in Yuncheng City. The average ozone formation potential (OFP) of VOCs was 162.88 µg·m-3, in which OVOCs had the highest contribution rate (45.37%); acetaldehyde, propionaldehyde, ethylene, isoprene, and toluene were the key active components; and industrial emission sources were the emission sources with the highest contribution rate. The average value of secondary organic aerosol formation potential (SOAp) of VOCs was 0.40 µg·m-3, in which the contribution rate of aromatic hydrocarbons was the highest (88.00%), and the solvent use source was the emission source with the highest contribution rate.

[Spatiotemporal Distribution and Seasonal Characteristics of Regional Transport of PM2.5 in Yuncheng City].

The spatiotemporal distribution characteristics of PM2.5 pollution in Yuncheng City were analyzed based on the ambient air quality monitoring data from March 2019 to February 2020, and the hybrid single-particle Lagrangian integrated trajectory (HYSPLIT) and cluster analysis were used to discuss the contamination transportation pathway and the distribution of potential pollution sources affecting PM2.5 in Yuncheng City in different seasons. The results showed that the PM2.5 concentration in Yuncheng was the highest in winter (111.24 µg·m-3) and the lowest in summer (30.02 µg·m-3). PM2.5/PM10 was greater than 0.6 in autumn and winter, indicating that the particulate matter pollution in Yuncheng City in autumn and winter was mainly fine particulate matter; spatially, the annual mean value of PM2.5 presented distribution characteristics that were high in the northern and central regions and low in the eastern and western regions. The high-value areas of PM2.5 were significantly and strongly correlated with SO2, NO2, and CO, indicating that local emissions contributed to the high values. The highest values in spring and winter were located in Hejin City (58.50 µg·m-3) and Jishan County (142.33 µg·m-3), and the highest values in summer and autumn were located in Pinglu County (36.92 µg·m-3) and the Salt Lake area (62.94 µg·m-3), respectively. Transportation analysis showed that spring and winter were mainly affected by air masses transporting northwestward, accounting for 58.69% and 55.77%. In summer, air masses were mainly eastward and southward, and in autumn, pollution was from short-distance transport air masses from southwestern Henan. The largest ratio was 32.89%; the potential source area was the smallest in spring and summer; it then began to grow in autumn, and it was the widest in the winter and had the highest contribution. The main source area in spring and summer was located in western and central Henan, and in autumn and winter, source areas were concentrated at the junction of Shaanxi, Shanxi, and Henan and west of Shaanxi.

Nitric Oxide Inhibits Al-Induced Programmed Cell Death in Root Tips of Peanut (Arachis hypogaea L.) by Affecting Physiological Properties of Antioxidants Systems and Cell Wall.

It has been reported that nitric oxide (NO) is a negative regulator of aluminum (Al)-induced programmed cell death (PCD) in peanut root tips. However, the inhibiting mechanism of NO on Al-induced PCD is unclear. In order to investigate the mechanism by which NO inhibits Al-induced PCD, the effects of co-treatment Al with the exogenous NO donor or the NO-specific scavenger on peanut root tips, the physiological properties of antioxidants systems and cell wall (CW) in root tip cells of NO inhibiting Al-induced PCD were studied with two peanut cultivars. The results showed that Al exposure induced endogenous NO accumulation, and endogenous NO burst increased antioxidant enzyme activity in response to Al stress. The addition of NO donor sodium nitroprusside (SNP) relieved Al-induced root elongation inhibition, cell death and Al adsorption in CW, as well as oxidative damage and ROS accumulation. Furthermore, co-treatment with the exogenous NO donor decreased MDA content, LOX activity and pectin methylesterase (PME) activity, increased xyloglucan endotransglucosylase (XET) activity and relative expression of the xyloglucan endotransglucosylase/hydrolase (XTH-32) gene. Taken together, exogenous NO alleviated Al-induced PCD by inhibiting Al adsorption in CW, enhancing antioxidant defense and reducing peroxidation of membrane lipids, alleviating the inhibition of Al on root elongation by maintaining the extensibility of CW, decreasing PME activity, and increasing XET activity and relative XTH-32 expression of CW.