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Genetic Network Inference: From Co-Expression Clustering to Reverse Engineering

Advances in molecular biological, analytical, and computational technologies are enabling us to systematically investigate the complex molecular processes underlying biological systems. In particular, using high-throughput gene expression assays, we are able to measure the output of the gene regulatory network. We aim here to review datamining and modeling approaches for conceptualizing and unraveling the functional relationships implicit in these datasets. Clustering of co-expression profiles allows us to infer shared regulatory inputs and functional pathways. We discuss various aspects of clustering, ranging from distance measures to clustering algorithms and multiple-duster memberships. More advanced analysis aims to infer causal connections between genes directly, i.e., who is regulating whom and how. We discuss several approaches to the problem of reverse engineering of genetic networks, from discrete Boolean networks, to continuous linear and non-linear models. We conclude that the combination of predictive modeling with systematic experimental verification will be required to gain a deeper insight into living organisms, therapeutic targeting, and bioengineering.

Dhaeseleer, Patrik

Acoustic space occupancy: Combining ecoacoustics and lidar to model biodiversity variation and detection bias across heterogeneous landscapes

There is global interest in quantifying changing biodiversity in human-modified landscapes. Ecoacoustics may offer a promising pathway for supporting multi-taxa monitoring, but its scalability has been hampered by the sonic complexity of biodiverse ecosystems and the imperfect detectability of animal-generated sounds. The acoustic signature of a habitat, or soundscape, contains information about multiple taxa and may circumvent species identification, but robust statistical technology for characterizing community-level attributes is lacking. Here, we present the Acoustic Space Occupancy Model, a flexible hierarchical framework designed to account for detection artifacts from acoustic surveys in order to model biologically relevant variation in acoustic space use among community assemblages. We illustrate its utility in a biologically and structurally diverse Amazon frontier forest landscape, a valuable test case for modeling biodiversity variation and acoustic attenuation from vegetation density. We use complementary airborne lidar data to capture aspects of 3D forest structure hypothesized to influence community composition and acoustic signal detection. Our novel analytic framework permitted us to model both the assembly and detectability of soundscapes using lidar-derived estimates of forest structure. Our empirical predictions were consistent with physical models of frequency-dependent attenuation, and we estimated that the probability of observing animal activity in the frequency channel most vulnerable to acoustic attenuation varied by over 60%, depending on vegetation density. There were also large differences in the biotic use of acoustic space predicted for intact and degraded forest habitats, with notable differences in the soundscape channels predominantly occupied by insects. This study advances the utility of ecoacoustics by providing a robust modeling framework for addressing detection bias from remote audio surveys while preserving the rich dimensionality of soundscape data, which may be critical for inferring biological patterns pertinent to multiple taxonomic groups in the tropics. Our methodology paves the way for greater integration of remotely sensed observations with high-throughput biodiversity data to help bring routine, multi-taxa monitoring to scale in dynamic and diverse landscapes.

Airborne lidar

Optimizing Single Nuclei Sequencing of Brain Samples From Space Flown Mice Across Age and Strain

The NASA GeneLab Sample Processing Laboratory offers high-throughput sequencing services to NASA-funded space biology researchers. Space biology studies have specific challenges such as low sample numbers, introducing susceptibility to batch effects from sample handling. These issues are compounded by complex protocols such as single-nuclei isolation and sequencing, which has recently become an attractive methodology for assessing the cellular diversity within spaceflight samples. High quality single-nuclei sequencing requires reproducible protocols to dissociate tissue and generate clean suspension of intact single nuclei. Producing single-nuclei suspension from brain tissue is particularly challenging due to cell type heterogeneity and the myelin sheath that carries over into the nuclei suspension as debris. Current procedures tend to be time consuming and sometimes include steps that can alter gene expression and create cell-type bias. Commercially available nuclei isolation kits, such as the 10X Genomics nuclei isolation kit, offers a streamlined way to process samples for nuclei isolation, thereby minimizing batch effects and enabling reproducibility. In this study, we report on the performance of the 10X Genomics nuclei isolation kit and Chromium Next GEM Single Cell Multiome ATAC + Gene Expression kit to generate sequencing libraries from space-flown mouse brain samples. Single nuclei sequencing was performed on frozen mouse brain tissue from two spaceflight missions, Rodent Research-10 (RR-10) and RR Reference Mission-2 (RRRM-2). RR-10 mice were female B6129SF2/J, euthanized at 18-19 weeks whereas RRRM-2 mice were female C57BL/6NTac, euthanized at 20 or 37 weeks. Sequencing data was processed using standard GeneLab data processing pipelines. We report evaluation of the performance of the 10X Genomics nuclei isolation kit for spaceflight samples from mouse brain, and evaluation of reproducibility across different mouse strains and age groups. We also report preliminary scientific results including cell type inference, cell clustering, and differentially expressed genes and pathways between spaceflight and ground control samples.

RR-10