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Packaging therapeutic RNAs for targeted treatment of breast cancer
Date:4/18/2012

is ensuring that the siRNAs get to where they need to go. "Naked siRNAs are cleared by the kidneys very quickly, and cells don't take them up readily," Lieberman explained. "Other delivery systems, like liposomes, tend to make siRNAs collect in the liver.

"The challenge is to deliver the siRNAs to the site of the tumor," she added, "without creating toxicity or inflammation in other portions of the body."

Lieberman and Song decided to adapt a targeting method they had originally developed for eliminating HIV-infected cells to transport siRNAs to breast tumors. First they bound siRNAs that would turn off a gene called PLK1 (an enzyme that promotes cell division) to protamine, a peptide that shrink-wraps DNA in sperm cells. Protamine was attached to an antibody fragment (called an ScFv) against HER2, creating a delivery package addressed directly to breast cancer cells with HER2 on their surface.

"The protamine stabilizes the siRNAs and protects them from being degraded by enzymes in the blood stream," Lieberman said. "And the ScFv antibody fragment makes sure that they only get delivered to and act in HER2-positive cells."

The team administered both naked and ScFv-protamine packaged siRNAs against PLK1 to mice with human breast tumors. While the naked siRNAs quickly accumulated in the kidneys and were excreted—their half life within the mice was a mere six minutes—Lieberman and Song found that the packaged ones were delivered directly to the HER2-positive tumor sites within the mice. There they were readily detectable for at least 72 hours.

Once in the tumors, the siRNAs were absorbed into tumor cells, where they turned off PLK1, retarding tumor growth and suppressing metastasis compared to untreated controls or those that received naked siRNAs.

The team found no evidence of inflammation or toxicity in the liver, kidneys, or bloodstream, indicating that the siRNAs were silencing PLK1
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SOURCE Boston Children's Hospital
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