Engineered bidirectional communication mediates a consensus in a microbial consortium
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Engineered bidirectional communication mediates a consensus in a microbial consortium. Presented by: Iman Mirrezaei. Outline. Introduction Problem definition Methods Experiments. Why am I still Sick . 170 Million American have Chronic disease What is Chronic bacterial disease?

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Engineered bidirectional communication mediates a consensus in a microbial consortium

Engineered bidirectional communication mediates a consensus in a microbial consortium

Presented by: ImanMirrezaei


Outline

Outline

  • Introduction

  • Problem definition

  • Methods

  • Experiments


Why am i still sick

Why am I still Sick

  • 170 Million American have Chronic disease

  • What is Chronic bacterial disease?

    • Infection of bones

    • Make people suffer and make people die

  • The real number are staggering

    • 550,000 deaths annually

    • 30x the number of Aids patients

    • Equal to the number of cancer death

    • $94 Billion in medical costs


Biofilm

Biofilm

  • A biofilm is any group of microorganisms

  • The ability to manipulate these films would enable controlled studies of microbial ecosystem dynamics

Staphylococcus aureusbiofilm


Biofilm formation

BioFilm formation

  • Reversible Adsorption of Bacteria (sec.)

  • Irreversible attachment of bacteria (sec.-min.)

  • Growth and division of bacteria (hrs.-days)

  • Exopolymer production and BioFilm formation (hrs.-days)

  • Attachment of other organism to BioFilm( days- months)


Biofilm relationship

BioFilm relationship

  • The BioFilm could maintain different relationship with the hosts:

    • Mutualism: both benefit

    • Commensalism: one benefit one unaffected

    • Parasitism: one benefit, one harmed

    • Competition: neither benefit

    • Neutralism: both unaffected


Biofilms properties

BioFilms properties

  • Difficult to be Removed and Killed

  • Function as a single organism


Microbial consortia form

Microbial consortia form

  • Microbial consortia form generate a function that none is capable of alone.

    • global food chain to human weight gain

  • A microbial consortium is two or more microbial groups living symbiotically


Problem definition

Problem definition

  • Two colocalized populations of Escherichia coli converse by exchanging lactone signals.

  • This consensus function can be considered a logical AND gate

  • The ability to manipulate and control these films

Escherichia Coli


Engineered circuits

Engineered circuits

  • Engineered circuits have been used to control the behavior of single cells and cell populations

  • Engineered communication

    • Broadcasting

    • Small-molecule signals


Experiments requirements

Experiments Requirements

  • Engineered circuits have been used to control the behavior of single cells and cell populations in both time and space

  • Escherichia coli Bacterium


The mcc signaling network

The MCC “Signaling Network”

  • Components of the LasI/LasR and RhlI/RhlR quorum sensing systems found in Pseudomonas aeruginosa, that forms a biofilm in the lungs of cystic fibrosis patients

cystic fibrosis patients


The mcc signaling network1

The MCC “Signaling Network”


Modeling to minimize crosstalk

Modeling to Minimize Crosstalk

  • Minimize densities for consensus activation, maximize densities for isolation activation

  • Positive feedback loop

  • Need C4HSL for LasI and 3OC12HSL; need 3OC12HSL for RhlI and C4HSL (minimize crosstalk signal)


Liquid culture

Liquid Culture

  • GFP( target gene) in both

  • Separate chambers with passage of small molecules

  • Responses over 100-fold greater with communication

Circuit A (with B)

Circuit B (with A)

Circuit B

(alone)

Circuit A (alone)


Solid culture

Solid Culture

  • Cell types embedded separately in solid medium

  • Media placed in contact

  • Fluorescence decreases with distance from interface

  • Circuit A slower growth


Engineered bidirectional communication mediates a consensus in a microbial consortium

Increasing fluorescence from increasing concentration of signaling molecule


Biofilms consortium

Biofilms (consortium)

  • Circuit A – green (yellow); Circuit B – red (cyan)

  • Grow together and display MCC function (≥6dys)

  • No significant fluorescence separately


Biofilm communication in nature

Biofilm Communication in Nature

  • Quorum sensing to coordinate biofilm formation

  • Biofilm disruption

  • Fundamental Applications

    • Integrate cells sensing different stimuli (temp, pH, ...)

    • More communication partners

    • Communication other than quorum


Applied applications

Applied Applications

  • Mixed-culture batch reactors require quorum of each

  • Disrupt pathogenic BioFilms

  • Enzyme-prodrug pair; inactive toxin fragments

  • Materials synthesis

  • Surveillance for environmental changes

    • Epidemiology, material degradation


References

References:

  • 1. Hooper LV, Midtvedt T, Gordon JI (2002) Annu Rev Nutr 22:283–307.

  • 2. Kato S, Haruta S, Cui ZJ, Ishii M, Igarashi Y (2005) Appl Environ Microbiol

    71:7099–7106.

  • 3. Kleessen B, Blaut M (2005) Br J Nutr 93(Suppl 1):S35–S40.

  • 4. Macfarlane S, Woodmansey EJ, Macfarlane GT (2005) Appl Environ Microbiol

    71:7483–7492.

  • 5. Mishra S, Jyot J, Kuhad RC, Lal B (2001) CurrMicrobiol 43:328–335.

  • 6. Becskei A, Serrano L (2000) Nature 405:590–593.

  • 7. Becskei A, Seraphin B, Serrano L (2001) EMBO J 20:2528–2535.

  • 8. Kobayashi H, Kaern M, Araki M, Chung K, Gardner TS, Cantor CR, Collins JJ (2004) Proc Natl Acad Sci USA 101:8414–8419.


Engineered bidirectional communication mediates a consensus in a microbial consortium

Thank you


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