Yinshan Fang

Co-culture with stellate ganglia (SG) promotes the growth of mouse SCLC

ICI treatment reduces the growth of SCLC

PKA inhibition (PKAi) reduces the growth of primary human SCLC cells

High levels of ADRB2 promote the growth of mouse SCLC

ADRB2 regulates the growth of human SCLC cells

Sympathetic and parasympathetic nerve fibers in mouse SCLC

Nucleotide modifications deviate nanopore sequencing readouts, therefore generating artifacts during the basecalling of sequence backbones. Here, we present a reference-guided, iterative approach to polish modification-disturbed basecalling results. We show that such an approach is uniquely suitable for training biomolecule-specific high-accuracy basecallers, by improving the basecalling of both artificially-synthesized and real-world molecules. With demonstrated efficacy and reliability, we exploit the approach to precisely basecall therapeutic RNAs consisting of artificial or natural modifications. We first analyzed vaccine mRNAs, which are artificially modified to promote stability and reduce immunogenicity. Specifically, we quantified the sequence purity and integrity, the two most important quality metrics to be controlled during mRNA vaccine production. We also analyzed BioRNAs, which are human tRNA-based carriers for therapeutic RNA interference (RNAi) agents. Specifically, we examined modification hotspots, which are naturally incorporated in vivo during BioRNA production and essential for therapeutic efficacy. Our analysis expands the scope of therapeutic RNA quality control, from the conventional sequence-level to the current modification status-level.

Tissue-resident immune cells such as innate lymphoid cells (ILC) are known to reside in the parenchymal compartments of tissues and modulate local immune protection. Here we use intravascular cell labeling, parabiosis and multiplex 3D imaging to identify a population of group 3 ILCs in mice that are present within the intravascular space of lung blood vessels (vILC3). vILC3s are distributed broadly in alveolar capillary beds from which inhaled pathogens enter the lung parenchyma. By contrast, conventional ILC3s in tissue parenchyma are enriched in lymphoid clusters in proximity to large veins. In a mouse model of pneumonia, Pseudomonas aeruginosa infection results in rapid vILC3 expansion and production of chemokines including CCL4. Blocking CCL4 in vivo attenuates neutrophil recruitment to the lung at the early stage of infection, resulting in prolonged inflammation and delayed bacterial clearance. Our findings thus define the intravascular space as a site of ILC residence in mice, and reveal a unique immune cell population that interfaces with tissue alarmins and the circulating immune system for timely host defense.