Team:HKUST-Hong Kong/Expectation

From 2012.igem.org

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           <p>Through assembly of the target binding  module, anti-tumor drug synthesis module and regulatory module, we would like to introduce our genetically engineered <i>Bacillus subtilis</i>, B. hercules, as an anti-colon-tumor agent to provide direct tumoricidal effect during cancer therapy. It is applied as an oral medicine which retains viability through gastrointestinal tract and executes anti-tumor activity when and only when it binds to colon cancer cell. </p>
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           <p>Through assembly of the target binding  module, anti-tumor drug synthesis module and regulatory module, we would introduce our genetically engineered <i>Bacillus subtilis</i>, B. hercules, as an anti-colon-tumor agent to provide direct tumoricidal effect during cancer therapy. It would be applied as an <u>oral medicine</u> which should retain viability in digestive tract and execute anti-tumor activity when and only when it binds to colon cancer cell. </p>
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<p>Driven by pVeg constitutive promoter,  RPMrel, the colon-tumor specific peptide will be displayed on the cell wall of  B. hercules under the facilitation of LytC cell wall displaying system before it is orally taken. When B. Hercules is taken orally by the patient, it is expected to reach colon in <strong>8 hours</strong> without retaining or colonizing in gastrointestinal tract. However, once it reaches colon, it will attach to colon tumor cells and colonize around them, waiting for signals to trigger the production and release of anti-tumor molecule, BMP2 to the local environment. <br />
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<p>Driven by pVeg constitutive promoter,  RPMrel, the colon-tumor specific peptide should be displayed on the cell wall of  B. hercules under the facilitation of LytC cell wall displaying system before oral ingestion. Once ingested, B. Hercules would be  expected to reach colon after <strong>8 hours</strong> without retaining in or colonizing the gastrointestinal tract. However, upon reaching the colon, it should hold up around colon tumor cells and colonize around them, and should await signals to produce anti-tumor molecule, BMP-2, which would be released to the local environment. <br />
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   Since the expression of BMP2 is controlled by xylose inducible promoter and no xylose is present in colon, xylose is taken orally or injected from anus to induce the  production of BMP2 when B. Hercules has successfully localized around colon tumor. The locally concentrated BMP2 is expected to suppress colon tumor growth and trigger the apoptosis of tumor while the low level BMP2 in non-tumor colon area introduces as little adverse effect as possible to normal colon tissues. <br />
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   The expression of BMP-2 would be controlled by xylose inducible promoter. Since normally no xylose is present in colon, xylose would be taken orally or injected from anus. At that moment, B. Hercules should has successfully localized around colon tumor, and the arriving xylose should induce the  production of BMP-2. The locally concentrated BMP-2 is expected to suppress colon tumor grow and trigger the apoptosis of the tumor. The BMP-2 level in colon should be low, and should have little adverse effect on normal colon tissues. <br />
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  Driven by the same promoter that regulates BMP2, autotoxin-encoding gene ydcE is simutaneously expressed and will gradually overwhelm the protection machinery by antitoxin ydcD. The overexpression of the autotoxin with BMP2 production will eventually cause the apoptosis of B. hurcules and prevent any adverse effect from BMP overdose.</p>
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The concentration of xylose might overshoot, which might result in excessive production of BMP-2. If that happened, the excess xylose would induce high level of expression of the toxin YdcE. Once the amount of YdcE breaches the protection  threshold from antitoxin YdcD, uninhibited YdcE will lead to the destruction of B. hercules. This would hopefully avoid the overdosing effect of xylose and the overproduction of BMP-2, and their subsequent adverse effect.</p>
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Revision as of 18:07, 24 September 2012

Team:HKUST-Hong Kong - 2012.igem.org

EXPECTATION

Through assembly of the target binding module, anti-tumor drug synthesis module and regulatory module, we would introduce our genetically engineered Bacillus subtilis, B. hercules, as an anti-colon-tumor agent to provide direct tumoricidal effect during cancer therapy. It would be applied as an oral medicine which should retain viability in digestive tract and execute anti-tumor activity when and only when it binds to colon cancer cell.

Driven by pVeg constitutive promoter, RPMrel, the colon-tumor specific peptide should be displayed on the cell wall of B. hercules under the facilitation of LytC cell wall displaying system before oral ingestion. Once ingested, B. Hercules would be expected to reach colon after 8 hours without retaining in or colonizing the gastrointestinal tract. However, upon reaching the colon, it should hold up around colon tumor cells and colonize around them, and should await signals to produce anti-tumor molecule, BMP-2, which would be released to the local environment.
The expression of BMP-2 would be controlled by xylose inducible promoter. Since normally no xylose is present in colon, xylose would be taken orally or injected from anus. At that moment, B. Hercules should has successfully localized around colon tumor, and the arriving xylose should induce the production of BMP-2. The locally concentrated BMP-2 is expected to suppress colon tumor grow and trigger the apoptosis of the tumor. The BMP-2 level in colon should be low, and should have little adverse effect on normal colon tissues.
The concentration of xylose might overshoot, which might result in excessive production of BMP-2. If that happened, the excess xylose would induce high level of expression of the toxin YdcE. Once the amount of YdcE breaches the protection threshold from antitoxin YdcD, uninhibited YdcE will lead to the destruction of B. hercules. This would hopefully avoid the overdosing effect of xylose and the overproduction of BMP-2, and their subsequent adverse effect.