Bacteria constantly release signal molecules If cell density is not big enough nothing happens ...

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Quorum Sensing and Biofilms

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Transcript of Bacteria constantly release signal molecules If cell density is not big enough nothing happens ...

Page 1: Bacteria constantly release signal molecules If cell density is not big enough nothing happens Threshold density all bacteria simultaneously.

Quorum Sensing and Biofilms

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Bacteria constantly release signal molecules If cell density is not big enough nothing

happens Threshold density all bacteria

simultaneously carry out gene expression “Quorum Sensing” from counting as to take

a vote before acting

Quorum Sensing Basics

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Acyl homoserine lactones (AHL molecules) Easy transport of molecules outside of cells

Gram Negative Bacteria

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Precursor Oligopeptides Must be exported via active transport

proteins

Gram Positive Bacteria

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Al-2 molecule is universal Produced by LuxS enzyme from dihydroxy

pentanedione substrate

Inter Species Communication

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Microorganisms form layer with proteins, DNA and long chain sugars

Often pathogenic Concern for health and food industry

◦ Spoiled food◦ Biofilms in cystis fibrosis

Quorum sensing plays big role in the high cell density communication and virulence

Biofilms

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Scientists are investigating varied bio-film formations and mutations that could help decrease formation

Looking at mutant luxS enzymes and other quorum sensing systems

Disrupt quorum sensingdisrupt biofilm formation and virulence

Biofilms Today

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Ep-CAM Gene Silencing in

Craniopharyngiomas

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Benign epithelial brain tumor common in children

Cannot spread but presses on vital structures

Can cause stunted growth, obesity, diabetes insipidus, and other morbidity

Hard to remove due to strong adherence to important structures

Often reoccurs

Background

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Ep-CAM gene expression is highly over expressed

Produces Ep-CAM glycoprotein adhesion molecule

Is linked to tumor genesis and metastasis Makes tumor strongly adherent

Ep-CAM Glycoprotein

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Craniopharyngioma

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Radiotherapy◦ Kills healthy cells◦ High risk of second cancer in children and adolescents

Competing Technologies

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Silencing Ep-CAM gene by inhibiting transcription through methylation of the promoter region

Design

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Inducing Bacteria

Aerotolerant E.Coli bacteria engineered to produce invasin protein and produce m5c DNA methyltransferase enzymes on signal

Calcification material

Ep-CAM glycoproteinReceptor

Receptor

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Invasin and m5c Production

m5c methyltransferase enzymes

Invasin proteins

Bacteria Produces

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B-1 Integrin receptor

Invading Tumor Cells

Tumor cell

Invasin Proteins

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Inside Cell

Ep-CAM promoter

M5c DNA methyltrasnferase enzymes

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Inhibition of RNA Polymerase Binding

RNA Polymerase

Ep-CAM Transcription

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Ep-CAM glycoprotein

Calcium deposits m5c methyltransferase production

0 0 0

0 1 0

1 0 0

1 1 1

AND Gate Truth Table

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Methylated CpG Islands on Promoter

Ep-CAM Expression

0 1

1 0

Inverter Logic Gate Truth Table

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No damage to healthy cells Easy to administer No radioactive material Helps surgeons remove tumor Decrease reoccurrence

Advantages

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Ep-CAM promoter region specific methylation

Laboratory precaution while working with cells with invasin

Precautions to keep bacteria from evolving and reproducing out of control

Possible Problems

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Testing on tissue culture and many other steps before reaching human testing

Similar system could be tweaked and developed for other cancers

Testing


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