Note: Complete set of data and code is available upon request

Interleukin-17 in Botryllus schlosseri

Group: Anthony De Tomaso Lab

Partner: Soham Ray, UCLA Environmental Engineering PhD student

Abstract

  Macrophages are immune cells with the ability to engulf cells and digest them by a process known as phagocytosis. While their roles in immune defense, homeostasis, cancer, and regeneration are well known, this project sought to understand the fundamental molecular/cellular mechanisms within a particular family of cytokines -- interleukin-17 (IL-17). The unique extracorporeal vasculature of the model organism, Botryllus schlosseri, was exploited to dissect the mechanisms of these cells during two opposing vascular remodeling events: Vascular regression and Vascular regeneration, using RNA/protein bioinformatics. We discovered through RNA sequencing data that IL-17 ligand expression was greatest during the early late stages of vascular regeneration, and early stages of vascular regression, whereas the receptors were consistently expressed. Additionally, phylogenetic analysis coupled with the genomic location of IL-17 revleaed that several IL-17 members are a result of gene duplication which is a precursor for the evolution of interleukins. This study helped to elucidate the ability of macrophages to transform into either inflammatory or anti-inflammatory under a given condition.

Botryllus schlosseri Blastogenetic Cycle

Botryllus schlosseri undergoes a week long blastogenetic cycle, regenerating its entire body de novo. It is a well choreographed dance of regenerating and regressing vasculature. We hypothesized that this process was aided by the immunoregulatory family of proteins, IL-17. Given the translucent nature of the extracorporeal vasculature, this 27 second long timeplapse captures the process.

Poster Presentation

Methods Used

• Phylogenetic Tree assembly using Maximum Likelihood
• Genomic Position and Direction of IL-17 candidate genes
• RNA sequencing during each distinct blastogenetic stage of marine tunicate (Stages A, B, C, D)
• Micro-injections of fluorescent bioparticles to simulate infections
• RT-qPCR to quantify expression of IL-17 ligands/receptors during simulated infection

Technologies

• R
• Python
• RNA-sequencing
• RT-qPCR
• Micro-injections
• Fluorescence microscopy

Contact

• Protein expression of target IL-17 receptors/ligands is a work in progress
• Feel free to contact me with any questions or if you are interested in contributing or collaborating!