Engineering Cell Function for Novel Therapeutics

The Bowles Lab works to engineer cell function by utilizing multiplex CRISPRa/CRISPRi systems and genome-wide screens to develop novel therapeutics for a broad range of disorders, including the musculoskeletal system, nervous system, and fibrosing disorders. As the first to investigate CRISPRi regulation in MSC/ASC populations, the lab has pioneered multiplex regulation and genome-wide screens for tissue engineering and gene therapy applications. The recent discovery of ZNF865 (BLST), a novel regulator of senescence and DNA damage, offers significant therapeutic potential for treating musculoskeletal disease, cancer, neurodegenerative disorders, and aging.

Digital diagram showing a central blue cell surrounded by representations of pH decrease, senescence, mitochondrial dysfunction with ROS, decreased ECM content, hypoxia, and inflammatory cytokines TNF-a, IL-1B, and IL-6. Digital diagram showing a central blue cell surrounded by representations of pH decrease, senescence, mitochondrial dysfunction with ROS, decreased ECM content, hypoxia, and inflammatory cytokines TNF-a, IL-1B, and IL-6.

Novel Therapeutics for Degenerative and Age-Related Diseases

Using a multitude of CRISPR-based tools including CRISPR activation (CRISPRa), CRISPR interference (CRISPRi) and genome wide CRISPR gene perturbation screens, the lab works to discover, study, and modulate the expression of genes that are involved in the progression of degenerative and age-related diseases. The historic focus of the lab has been on the intervertebral disc; however,  the scope of our research has broadened to include additional musculoskeletal conditions, neurodegenerative diseases, and cancer. 

Flowchart showing the biological pathway of ZNF865. High expression leads to healthy IVD homeostasis, while low expression leads to cellular senescence, resulting in inflammation, tissue catabolism, immune cell recruitment, and nociceptor sensitization. Flowchart showing the biological pathway of ZNF865. High expression leads to healthy IVD homeostasis, while low expression leads to cellular senescence, resulting in inflammation, tissue catabolism, immune cell recruitment, and nociceptor sensitization.

ZNF865 (BLST)

The gene ZNF865 (BLST) is a novel modulator of senescence and DNA repair. When the Bowles lab began studying this gene, there were no publications on its function, and ongoing projects are actively working to elucidate it's involvement in numerous cellular pathways. Preliminary studies have shown that this gene can prevent and/or rescue cells from entering a senescent state and is heavily involved in the DNA damage response pathway. In addition to deducing the underlying biological processes regulated by this gene, we are also exploring its potential translatability as a therapeutic for musculoskeletal disease, cancer, neurodegenerative disorders, and other aging-related diseases.

A vector diagram showing a green dCas9 protein, yellow sgRNA, and an activator complex consisting of VP64, p65, and Rta, interacting with a DNA strand to trigger gene expression via RNA Pol II. A vector diagram showing a green dCas9 protein, yellow sgRNA, and an activator complex consisting of VP64, p65, and Rta, interacting with a DNA strand to trigger gene expression via RNA Pol II.

Multiplex CRISPR Gene Modulation

Using advancements in CRISPR activation and interference technology, the lab works to develop multiplex therapeutics that target several genes involved in a pathogenic state at once. In the musculoskeletal space, this includes the development of multiplex vectors targeting collagen and aggrecan production. Additionally, we have worked to modulate the expression of multiple inflammatory cytokines involved in age-related degenerative conditions.