Toward an Integrated Computational Framework for Metagenomics: From Sequence Alignment to Automated Knowledge Discovery

Metagenomic sequencing has become a central paradigm for studying complex microbial communities and their interactions with the host, with emerging applications in clinical prediction and disease modeling. In this work, we first investigate two representative application scenarios: predicting immune checkpoint inhibitor response in non-small cell lung cancer using gut microbial signatures, and characterizing host–microbiome interactions in neonatal systems. The proposed reference-free neural network captures both compositional and functional signals without reliance on reference genomes, while the neonatal study demonstrates how environmental and genetic factors reshape microbial communities and how probiotic intervention can mitigate pathogen-induced immune activation.

These studies highlight both the promise and the inherent difficulty of metagenomic analysis: transforming raw sequencing data into clinically actionable insights remains an algorithmically fragmented and computationally intensive process. This challenge arises from two key limitations: the lack of a unified algorithmic foundation for sequence alignment and the absence of systematic approaches for selecting and organizing analytical tools. Motivated by these challenges, we present a unified computational framework for metagenomic analysis that integrates complementary algorithmic and systems-level solutions.

First, to resolve fragmentation at the alignment level, we develop the Versatile Alignment Toolkit (VAT), a unified algorithmic system for biological sequence alignment across diverse applications. VAT introduces an asymmetric multi-view k-mer indexing scheme that integrates multiple seeding strategies within a single architecture and enables dynamic seed-length adjustment via longest common prefix (LCP)–based inference without re-indexing. A flexible seed-chaining mechanism further supports diverse alignment scenarios, including collinear, rearranged, and split alignments. Combined with a hardware-efficient in-register bitonic sorting algorithm and dynamic index-loading strategy, VAT achieves high efficiency and broad applicability across read mapping, homology search, and whole-genome alignment. Second, to address the challenge of tool selection and pipeline construction, we develop SNAIL, a natural language processing system for automated recognition of bioinformatics tools from large-scale and rapidly growing scientific literature. By integrating XGBoost and Transformer-based models such as SciBERT, SNAIL enables structured extraction of analytical tools and supports automated, reproducible pipeline construction.

Together, this work establishes a unified framework that is grounded in real-world applications and addresses key bottlenecks in metagenomic analysis, enabling more efficient, scalable, and clinically actionable workflows.