Detection and treatment of critical illnesses using oligonucleotides

Abstract

Sepsis is among the most prevalent diagnosed critical illnesses in the United States today. Although advances have reduced the overall morbidity and mortality associated with this illness, the enormous number of deaths associated with it shows a need for improved diagnostic and therapeutic options. Our laboratory has utilized RNA based technologies to aid in the treatment of histone induced multiple organ dysfunction syndrome seen in sepsis.Histones are proteins found in the nucleus of every cell in our body and have been shown to be released during sepsis. Such release induces damage to other cells, causing a feed forward cycle that results in organ failure and death. Several therapeutics have been utilized to neutralize histones but have shown considerable toxicity. This thesis describes the generation of single stranded RNA aptamers to bind and neutralize histone mediate damage without unwanted toxicity. We demonstrate that our aptamers selectively bind to histones but not serum proteins. In addition, we establish that our aptamers can neutralize all histone mediated cellular response in vitro and in vivo. Finally, we determined that our aptamers are able inhibit the histone feed forward cycle in a temporal fashion in our murine model of multiple organ dysfunction. This novel therapeutic demonstrates the selectivity and effectiveness needed to inhibit histones in several critical illnesses. iv PUBLIC ABSTRACTThe most prevalent diagnosed critical illness in the western hemisphere is sepsis.Despite recent advances in treatments, the number of deaths associated with sepsis continues to grow, resulting in the need for improved diagnostic and therapeutic options.Our laboratory has utilized RNA based technologies to aid in the treatment and detection of the critical illness stated above.During a critical illness such as sepsis, cells are damaged causing the release of proteins from the nucleus (i.e. histones) resulting in further cellular damage that leads to organ failure and death. To date, one FDA approved therapeutic has been utilized to neutralize histones but resulted in considerable toxicity in the patients. In this thesis we describe a novel therapeutic (single-stranded RNA aptamers) to bind and neutralize histone mediated damage without the toxic side-effects. This novel therapeutic demonstrates the selectivity and efficacy needed to inhibit histones in several critical illnesses.