The MBD5/6 complex is recruited to methylated DNA to promote the silencing of genes and transposable elements (TEs). As more MBD5/6 is recruited to a region in the genome, multiple genes and TEs are silenced successfully, shutting off their transcription. One of the complex’s components is heat shock proteins (HSPs), which oligomerize with each other to promote the accumulation of MBD6 proteins at specific loci and achieve DNA silencing. Such as how heat shock proteins can behave as chaperones in A. thaliana, it was sought out to determine if human heat shock proteins (hHSPs) would act similarly. Chimeric sequences of MBD6-hHSP1, MBD6-hHSP3, MBD6-hHSP5, and MBD6-hHSP8 with a fluorescent protein were created via molecular cloning. To demonstrate hHSP chaperoning abilities, the four MBD6-hHSP sequences with a fluorescent protein were genomically inserted into different backgrounds of A. thaliana. Microscope visualization confirmed that all hHSPs, except for hHSP8, showed an accumulation of MBD6 at bright foci, indicating hHSP oligomerization and DNA silencing. The hHSPs that generated bright foci can be used with the Suntag system, which is a system that grants the ability to direct proteins to a desired destination in the genome. From previous studies, the Suntag system and HSPs were able to direct the concentration of MBD6 at specific loci only when HSPs were present. Determining if hHSPs could behave similarly to HSPs with the Suntag system is significant to discover since it can grant the user the ability to direct silencing proteins to a desired genomic site.

In lung disease, cardiac dysfunction is known to occur, but the mechanism underlying cardiac dysfunction remains uncertain. One model is that this occurrence results from cross-talk in the stellate ganglion (SG). In this study, our aim is to better characterize the anatomic and neurochemical profile of heart- and lung-projecting stellate ganglion neurons in normal physiology, while also describing the remodeling of these neurons in pulmonary fibrosis (PF) using a mouse model of bleomycin-induced PF. Our hypothesis posits that mice with PF will undergo morphological and neurochemical remodeling of both heart- and lung-projecting neurons within the SG. The fraction of total lung-projecting SG neurons immunoreactive for neuropeptide Y (NPY) was 59% in the control (saline) group (n=9) and 79% in the bleomycin group (n=12), p=0.0012. In-situ hybridization was performed and preliminary data is demonstrating similar trends of ribonucleic acid expression for NPY within the SG in PF (p=0.171). Additionally, the immunoreactivity of choline acetyltransferase and vasoactive intestinal peptide was investigated for heart- and lung-projecting SG neurons. Most recently, retrograde neural tracing using adeno-associated viruses has been optimized in preparation for single-cell RNA sequencing to study the transcriptome of heart- and lung-projecting SG neurons. In conclusion, our findings demonstrate remodeling of heart- and lung-projecting SG neurons in response to bleomycin-induced PF. Neurochemical changes that occur within the stellate ganglion may elucidate novel therapeutics for PF, and the craniomedial region of the SG may serve as a neuroanatomical basis for cardiopulmonary crosstalk.