Blood microbial DNA signature differentiates hepatocellular carcinoma from metastatic lesions

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The microbiome influences cancer development, progression, and therapy response 46 across various malignancies [1, 2, 3, 4, 5].The composition and diversity of microbial 47 communities within various tissues, influence cancer development, progression, and therapy 48 response.Understanding specific microbial signatures associated with cancers, termed the 49 oncobiome, is crucial for advancing diagnostic and therapeutic strategies.50 Analysis of microbial reads from whole-genome sequencing data of blood and tissue 51 samples from The Cancer Genome Atlas (TCGA) identified microbial communities 52 distinguishing hepatocellular carcinoma (HCC) from non-cancerous tissue and other cancers 53 [2].These findings are robust across multiple rigorous analysis pipelines.Additionally, 54 circulating microbiome DNA has shown promise as a biomarker for early lung cancer diagnosis 55 and recurrence, suggesting the viability of blood-based microbial detection platforms [2].56 Furthermore, recent research on metastatic tissues indicated microbial communities align more 57 closely with the metastatic site rather than the primary cancer origin, though this was not 58 evaluated in blood [1].59 However, significant challenges remain for microbial cell-free DNA (cfDNA) to effectively 60 distinguish primary from secondary liver tumors.Previous studies have not demonstrated if 61 microbial cfDNA differentiates between HCC and metastatic liver tumors.Additionally, hepatitis 62 B virus (HBV) DNA contamination limited the ability to clearly separate HCC from non-HCC 63 samples.Thus, it remains uncertain whether liver cancers share a common microbial cfDNA 64 profile or if primary liver cancers possess a distinct signature.In this study, we apply shotgun 65 metagenomics to cfDNA to identify unique blood-based bacterial signatures that differentiate 66 HCC from metastatic colorectal cancer (mCRC).67 We assessed the metagenomics profile of tumor, adjacent tissue, and plasma samples 68 from participants diagnosed with HCC (n = 16) or mCRC (n = 11) (Supplementary Table 1).

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