Queenie chan

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20.109(F11): Synthesizing Hyaluronic Acid-Paclitaxel Conjugates using Viruses

Hannah Johnsen & Queenie Chan

Hyaluronic acid (HA) is a small molecule can be used as a nano-carrier for a cancer therapeutic. HA binds to CD44 receptors, which are commonly overexpressed on cancer cells, and once bound, are internalized into the cell.

References:

http://www.sciencedirect.com/science/article/pii/S0927776511006138

http://www.cancer.gov/cancertopics/druginfo/paclitaxel

http://www.nature.com/nnano/journal/v6/n6/full/nnano.2011.50.html

Project Overview: 1) target & amplification 2) image!

Background: HA: - internalized by cell via CD44 receptors, which are commonly overexpressed on cancer cells → can target cancer cells

IL1A increases expression of CD44 http://www.jbc.org/content/270/46/27734.full

Chelated Gd(III) as an imaging contrasting agent for MRI http://www.medicinenet.com/mri_scan/article.htm - can be used in cancer cells http://www.diagnosticimaging.com/dimag/legacy/bodymri/hires.html http://www.insideradiology.com.au/pages/view.php?T_id=38 - “complex molec” = Gd(III) with carrier (chelating agent) http://www.ncbi.nlm.nih.gov/pubmed/9680558 - how much to use ( 0.1 mmol/kg for large veins) http://www.ncbi.nlm.nih.gov/books/NBK23314/ = Gd-DTPA

How to chemically conjugate HA to stuff: http://pubs.acs.org/doi/full/10.1021/bc8000485 utilize carboxylic and hyrodxyl groups on HA to conjugate whatever we want


Research Problem & Goals: image cancer better in in vitro tissue culture amplify signal → better detection for earlier stuffs


The Project Details & Methods: - make sure HA conjugated with IL1A doesn't alter protein structure

I. HA conjugated with interleukin-1α (IL1A) - make sure HA-IL1A still induces expression of CD44 (follow same procedure as http://www.jimmunol.org/content/162/8/4920.full.pdf) - check to see that can be internalized by cells and into nucleus

II. HA conjugated with Gd-DTPA - injected through blood - toxicity tests, remove cells after treating with Gd(III) to see if they grow and are still alive - check to see that cancer cells are targeted - test imaging with cancerous cell and noncancerous cells - how much is necessary for imaging? - base off of 0.1 mmol/kg body mass, also test with cancer/noncancer - what percent of stuff is internalized? --> how much needed to be injected

- check that the two-step process targets the same cells - fluorescently tag IL1A and then add Gd-DTPA, see if same spots show

Predicted Outcomes: - see lots of signal (light space), maybe blobs, if cancer cells present - if nothing, not as light

Needed Resources: - HA - IL1A - chelated Gd(III) - necessary reagents to conjugate the above

- necessary materials for HA-IL1A checking

- cells - chelated Gd(III)

- fluorescently tagged IL1A - chelated Gd(III)

- (for final finished product) tissue sample (from mole or tumors or other potentially cancerous cells)


Future tests: - targeting cancer cells in people for earlier detection? - check for immune response (in vivo studies)