What’s Our Focus?

----Intestinal S.aureus Colonization

Staphylococcus aureus is a major human pathogen that contributes to a variety of lethal diseases, causing a large burden of morbidity and mortality worldwide. Due to its great infectivity and the abuse of antibiotics, many strains have developed resistance to an array of antibiotics, in particular, methicillin resistant S. aureus (MRSA), which already becomes an acute clinical problem.

Despite the well-established risk factor of its colonization in human nose, the gastrointestinal colonization by Staphylococcus aureus is still ill-defined. The average reported detection rate of intestinal carriage is 20% for normal strains and 9% for MRSA. The high risk population is suggested to be infants, hospitalized patients with inflammatory bowel disease (IBD), patients with a history of MRSA colonization or infection, and those with poor immune system status.

Apart from severe diarrhea, fever and dehydration, it is suggested recently that transient or persistent intestinal S. aureus colonization may cause Pseudomembranous colitis, induce specific systemic immune resonses and even alter the overall structure of the human colonic microbiota and the microbial metabolic profiles. Although it’s not been thoroughly studied yet, we believe that the potential clinical impacts cannot be underestimated.

What’s Our Aim and Why?

On these concerns:

1. The lack of clinical experience when dealing with this rare symptom;
2. The difficulties to prevent and fully eliminate intestinal S. aureus colonization;
3. Abuse of antibiotics and its side effects.

Aim to:

1. Design a novel therapy to combat with, and even prevent the intestinal S. aureus colonization within a short time after infection, thus reducing the use of antibiotics and restraining the development and transmission of MRSA.
2. 2. Introduce a new concept medicine that is safe, effective, and has the potential to be industrialized and put into drug market (which needs approval of China's State Food and Drug Administration).

How We Make it Happen?

--Probiotic Therapy

We genetically engineered Lactococcus lactis to become our drug carrier (“Grenadier Guards”), which will become active when arriving human intestine after digesting.

With a copy of the quorum sensing system from S. aureus, they will quickly sense the presence of the pathogen cells that are above the threshold concentration and have the mobility to move near the colony. Simultaneously, the synthesis of antimicrobial peptides (“Grenades”) will begin. Osmoregulation and tandem repeating are applied here to control the synthesis of our AMPs and allow them to reach a relatively high quantity level. After AMPs accumulation, our Lactococcus lactis will go through an endolysis procedure to release the synthesized peptides to attack the biofilm of S. aureus. All of these metabolism should proceed within a short time, so when combined with its large quantity and proper anti-diarrhea drugs, the symptom is likely to be reduced and even eliminated.

Why using probiotic as a carrier?

1. Lactic acid bacteria (LAB) are one of the potential vehicles for drug delivery in the gastrointestinal tract. They are bile-resistant and have the ability to survive passage to the GI tract. More importantly, many of them are generally recognized as safe (GRAS) for human and animal consumption. So here we aim to use L. lactis as the carrier, being one of the most amenable expression cell factories for heterologous protein secretion.
2. Research has shown that certain lactic acid bacteria, mainly commercial probiotics, e.g. Lactococcus lactis, were able to reduce adhesion and viability of adherent S.aureus. (doi: 10.1099/mic.0.28522-0)
3. As a commercial probiotic, genetically-engineered L. lactis has the potential to be widely applied to dairy products, which makes the market even broader.

Why choosing antimicrobial peptides?

1. AMPs play a direct antimicrobial and mediator function and provide the initial host defense mechanism against invading pathogens, and they are found to have potential to overcome bacterial resistance.
2. Here we combined three AMPs to enhance the inhibiting effect: LL-37, GF-17 and grammistin.

What is the drug delivery method?

Enteric coated tablet: Protect the engineered probiotics from partial degradation in the acidic environment of the stomach, and release the drugs when it reaches intestine.

What Are the Impacts?

1. Clinical

Combined with anti-diarrhoea drug, our probiotic therapy can quickly detect and eliminate the adherent S.aureus, thus reliving the patients from suffering.

2. Daily

By applying the engineered probiotics into daily dairy products, the public can get easily access to the drugs, which may results in a prevention of potential S.aureus adherence.