Team:UFMG Brazil/ProblemAndSolution
Project
Lab Work
Modeling
Practices
Synenergene
Team
What is the problem?
Macrophage-mediated Inflammatory Disorders
Mediators of inflammation induced by macrophages are critical for a variety of human inflammatory disorders (Jou et al., 2013), such as Rheumatoid Arthritis and Gout. Once activated macrophages actively secrete and cause an imbalance of cytokines, chemokines, and mediators of inflammation (Jou et al., 2013).
Gout
Gout is a painful and potentially disabling form of arthritis. The first symptoms are usually intense episodes of painful swelling in single joints, most often in the feet, especially the big toe, which turns red and warm. 50% of first episodes occur in the big toe, but any joint can be involved. The diagnosis of gout can be difficult, and often treatment plans must be adapted to each person, including nutritional education and use of drugs (Schumacher, 2015).
Gout affects the quality of life due to intermittent attacks and the potential of becoming chronic (Schumacher, 2015). This disease occurs in about 4% of American adults (about 6 million men and 2 million women; Arthritis Foundation, 2015).
This disease occurs when excess uric acid (a normal waste product) collects in the body, and urate crystals deposit in the joints. Certain foods (such as shellfish and red meats, alcohol in excess, sugary drinks and foods that are high in fructose) and drugs (such as low-dose aspirin, certain diuretics, and immunosuppressants used in organ transplants) may raise uric acid levels and lead to gout attacks (Schumacher, 2015).
Over time, increased uric acid levels in the blood may lead to deposits of urate crystals in and around the joints. These crystals can attract white blood cells, leading to severe, painful gout attacks and chronic arthritis. Uric acid also can deposit in the urinary tract, causing kidney stones (Schumacher, 2015).
Rheumatoid Arthritis (RA)
Rheumatoid arthritis is a chronic autoimmune disease that leads to deformity and destruction of joints by bone and cartilage erosion. It is estimated that 1% to 3% of the world population is affected, being most common in developing countries. This disease affects about two to three times more women, aged between 30-50 years, with peak incidence in the fifth decade of life (Alamanos & Drosos, 2005; Mota et al., 2012). In Brazil, the prevalence of rheumatoid arthritis in adults ranges from 0.2% to 1%, corresponding to an estimated 1.3 million people affected (Alamanos et al., 2006; Ferraz & Ciconelli, 2003; Woolf & Pfleger, 2003; Laurindo et al., 2002; ACR Clinical Guidelins, 1996; Symmons et al., 1994; Marques Neto et al., 1993).
In 2007, RA was the fourth largest budgetary impact disease, consuming 10.4% of the resources (Carias et al., 2011). Studies have shown that the biological disease-modifying drugs have ongoing high cost, and the annual cost of treatment per patient estimated between € 13,500 (US$ 15,296.01) and € 15,000 (US$ 16,995.56; Hoebert et al., 2012). In Brazil, a 48-month course of treatment with conventional regimen followed by biological costs R$ 1,701,286.76 (US$ 438803,94; Monteiro & Zanini, 2008), and the cost of six months of treatment with infliximab is R$ 19,698.00 (US$ 5,080.60; Venson et al., 2011).
The mortality rate of patients with RA is higher than in normal population, and life expectancy decreases according to the severity of the disease and when it started (http://www.artritereumatoide.com.br/artrite-reumatoide/#epidemiologia).
Inflammatory macrophages are central cells to the pathophysiology of disease due to their prominent numbers in the inflamed synovial membrane and at cartilage-pannus junction, their clear activation status and their response to successful anti-rheumatic treatment.
There is no cure for RA and the drugs usually used for its treatment are anti-inflammatory compounds with oral and/or parenteral administration. Because of this, the typically prescribed medications potentially have serious systemic side effects. Besides, biological characteristics of the joints make the drug delivery difficult when targeting sites, compromising the efficacy of the treatment.
IFN-β
The type I interferons (IFNs), IFN-β and various IFN-αs, are pleiotropic cytokines acting on a range of cell types and eliciting a diverse range of responses. For many years, IFN-β was thought to be a potential agent for the treatment of a variety of immune-mediated diseases (Vervoordeldonk et al., 2009). This molecule has N-glycosylation sites that increase stability and activity of molecule (Sommereyns & Michiels, 2006).
IFN-β has clear anti-inflammatory properties (Tak, 2004) and plays an important role in bone homeostasis (Takayanagi et al., 2002). Also, IFN-β has the ability to reduce the secretion of proinflammatory mediators, such as interleukin (IL)6, tumour necrosis factor (TNF)α, matrix metalloproteinases (MMPs) and prostaglandin E2, which are key players in the pathogenesis of RA (Coclet-Ninin et al., 1997; Smeets et al., 2000). In addition, IFN-β has antiangiogenic properties (McCarty et al., 2002), which could boost therapeutic effect in RA (Gerlag et al., 2001).
The notion that IFN-β treatment may reduce arthritis is supported by animal experiments. Several studies examining the effect of IFN-β in collagen-induced arthritis (CIA) in mice have been published (Treschow et al., 2005; van Holten et al., 2004; Triantaphyllopoulos et al., 1999; Tak et al., 1999).
Treatment of RA patients with IFN-β has been unsuccessful so far, presumably as a result of pharmacokinetic issues (van Holten et al., 2005). Novel approaches leading to constitutive IFN-β production at the site of inflammation may be required to induce clinical efficacy in patients.
Lack of better chassis
Nowadays, Synthetic Biology studies are limited by a few numbers of chassis, such as bacteria (Escherichia coli and Bacillus subtilis) and yeast (Saccharomyces cerevisiae).
One of the major problems of drug therapies are the side effects: other non-target tissues can be affected, causing discomfort or even causing the appearance of another syndrome. A treatment directed at the diseased location without affecting surrounding tissue, may be more effective and cause side effects of lower intensity and less severe.
A problem-solving approach
A brand new chassis!
The protozoan Leishmania is a good new chassi:
1) It has the ability to infect macrophages (Selvapandiyan et al., 2004);
2) It has the ability to express proteins with proper post-translational modifications, such as glycosylation and disulfide bonds;
3) It has the ability to deliver those proteins inside macrophages.
Leishmania: relax, it is biosafe!
The visceral Leishmania donovani strain with the virulence factor Centrin-1 deleted (LdCen1) has very low risks of contamination, and its virulence reduction has been evaluated and confirmed with different animal models, such as mice (Selvapandiyan et al., 2006), hamsters (Selvapandiyan et al., 2009), and dogs (Fiuza et al., 2014).
The genetically targeted and defined attenuation reduces the risk of reversal to virulence, a concern generally raised for attenuated organisms that are created by random genomic mutations.
LdCen1 deletion specifically attenuated the amastigote stage of the parasite that replicates inside macrophages, and has no effect on the growth of the promastigote form (Selvapadiyan et al., 2012).
In addition, we propose a kill switch system based on the genetic control of enzyme 3'-nucleotidase/nuclease, which is crucial for the purine rescuing on the promastigote form of Leishmania, the one which is spread by the sandflies, aiming to keep the modified parasite from spreading. The enzyme's synthesis would be regulated by a Tetracycline repressor and so the Leishmania would be cultivated in the presence of Tetracycline.
Our proposal
We propose the use of an optimized visceral Leishmania strain to direct IFN-β to specific macrophages associated to to joint inflammatory diseases.
References
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