 from I.T. Rootkey that is set in at the fourth house of Himalayas and it is in north of India and it is actually the oldest technical college in India which was set up in 1853 and at that time it was called Thompson College of Engineering. Anyways, this summer I have been working with Dr. Webukai at the UW Carbon Cancer Center and I have been working on molecular energy to target tumor mangyogenesis and we have used antibodies as well as protein nanoparticles to do the same. Now, cancer as we know is the second leading cause of death in USA and the major problem that accompanies its treatment is its late detection. By the time it is detected it is already so way ahead into, I mean it is like way ahead into the disease so it is here becomes impossible. So, this is where molecular imaging can help us. Molecular imaging is the visualization, characterization and measurement of a biological process either at molecular or cellular level in living systems. And these imaging modalities these include positron emission primographies and optical imaging which includes fluorescence, vial fluorescence and magnetic resonance imaging etc. Now, to improve upon these imaging modalities either in terms of improving upon the imaging levels or just changing the imaging levels totally we can use nanotechnology which will help us in early detection as well as in accurate treatment of accurate diagnosis and treatment of cancer or even other diseases. And I think the place of molecular imaging and therapy and nanotechnology the Kailat works. Now, these are some of the small handle molecular imaging modalities that are available which includes optical as I said before MRI, PET, CT that is computed tomography ultrasound imaging and spread and the University of Wisconsin has 4 out of 6 of these which have been used for research. Now, the aspect of cancer that that microjet that microjet would be targeting is angiogenesis which is formation of new blood vessels and this is very important for initiation and progression of cancer. One of the proteins that are expressed mainly in the vascular endothelial cells during angiogenesis in proliferating tumors is CD105 and this is a transmembrane protein. So, this would be our target. The targeting ligand that we would be using in our project is TRC105 which is the monopermal antibody and this binds to both human and urine CD105. And in fact it is about to step into second phase trials in USA, clinical trials in USA and because PET is such a great imaging modality in fact it is one of the primary imaging modalities for tumor detection and due to its excellent sensitivity tumor penetration and its ability to be at the images of PET can be accurately quantified. So, we will be using PET that is positron emission tomography as one of the major imaging modalities for microjet. Now, generally like PET as of now in our clinical oncology it uses fluorideoxy glucose which is a which is the glucose molecule which is bound to fluorine, a radiolabin fluorine as you can see over here and this radio tracer is now injected into the body intravenously and where the this fluorine the PK-Soli emits positron, positrons which annihilate with electrons and release gamma rays which can be detected and this is how a tumor is imaged tumor or in fact any disease can be imaged. But the problem is that this FDG it is based on fact that it would be incorporated in glucose metabolizing regions and cancer as cancer has been shown to be a highly active metabolic tissue for glucose and FDG is not cancer specific. So, this can have high updates in normal organs which normally metabolize glucose very like at a very high rate for example, brain, heart and muscle and also in some conditions like inflammatory now conditions or some infectious diseases or after surgery the tissues in those regions they show a high update of glucose. So, this might result in false positives. So, that means that so we need some imaging modality or sorry some imaging tracer which would be very specific to tumor. So, this is where we use antibodies. Now, why do we bring in graphene at all which is nanoparticle? The thing is that nanoparticles they show they have a large surface area onto which several imaging labels can be conjugated. So, this would give a high signal dramatic signal amplification also we can attach different targeting ligands onto the nanoparticles. And so this would increase the binding affinity and specificity of the constructs and also nanoparticles have been shown to bypass biological barriers even the blood brain barriers in some cases. So, the targeting efficiency is enhanced and in future what can be done or is postulated is that a single nanoparticle can act as a substrate for holding the imaging tracers the ligands as well as the drops. So, that you can have simultaneous imaging diagnosis and cure and treatment of the disease. So, if that happens that would be like from some people who have read people's nanotechnology can is actually the only answer probably now to effectively deal with cancer. So, with this overview I would like to introduce you to my projects. The first one is we will try to target vascular tumor vascular using functionalized looking outside nano streets. And the second project would deal with developing an antibody which dual modality agent for a pet and optical imaging in breast cancer of 41 cells of breast cancer. So, moving on with my first project we have this protein outside nano sheet on to which we would conjugate the TRC-105 which is the antibody and on to which we would be conjugating a radio tracer using this is NOTA actually a collator for radio nucleates and so I will move on with my project. So, first of all with groupine what is groupine? Groupine is actually a monolayer of carbon atoms it is a two dimensional sheet in which carbon atoms are like take the form of hexagons and this is a very amenable nanomaterial because it can be easily wrapped up into nanomaterials or can be rolled into nanobus as you can see in this animation here and it can be styled on one or these can be styled on top of each other to form the fight and actually it has been very much in use since it won the Nobel Prize in 2010 when it was discovered and but it has not really been used in biomedical sciences as such it has mainly found uses in electronics. So, it is very easy for nanomaterial to work with because just like apart from the high surface area which all the nanomaterials provide it has also been shown that groupine is as the high believer stability it chose low toxicity and it is very easy to functionalize because it has a plane sheet like structure so it would be very easy to attach things on to it. Now this is my workflow so what we did was that we took groupine of side nano sheet which was around 10 to 15 nanomaterials in dimensions and we conjugated NOTA which is our biopunctional calculator for radio nucleolucreants on to it then we attached the antibody which would be our targeting ligand on to it. Now these constructs were then tested in mid-flow in VUX and by microscopy studies and facts analysis and after the successful like we got some good results with it and so we now in we like we did copper 64 which is the radio nucleolucreants for PET imaging on to it and these constructs were then intra-venous injected into breast tumor bearing mice and and we image them at different time points from 3 hours on to 48 hours and then the mice were sacrificed and biodistribution was carried out using gamma counter to validate the PET data. So, this is the first step where we conjugate NOTA on to dripping of side. NOTA is basically a trihazor, cyclononene, trihecetic acid, this is a macro cycle and the compound of NOTA that we use is actually benzyl isothinate derivative isothiocinate derivative of NOTA so that it would help us in like you know attaching it to dripping of side nano sheets. So, these dripping of side nano sheets are also functionalized with PET, PET would help us in to with help would help to act as a spacer between dripping and NOTA and similarly we attach the antibody to dripping of side using PET again and then we carried out facts and fluorescence microscopy studies the results I would not be able to show them to you, but so then we carried on with copper labeling and this NOTA here it so when we incubate with copper at the particular pH we see that the copper atom it goes and fits inside the cavity formed by the bridges or carbon bridges and the nitrogen atoms and these ones that you can see over here these are actually coordinate bonds so the bonds are pretty strong and so this is how we label it with the protein conjugates and now these conjugates are injected into a velocity in mice and as we can see these are the PET immigrants at 3R time point and 48R time point and the control would be just a plain graphene oxide nano sheet without any targeting antibody that is TRC105 and we can see a very good very good localization to the tumor in case of the conjugates but in case of graphene oxide we can hardly see any tumor uptake so this shows that the graphene oxide and the graphene oxide and TRC105 conjugates have a very high tumor like comparatively high tumor uptake and with time at 48 hours you can see there is a diminishing of signals so that is probably because of one copper 64 it has a definite half life of about 12.8 hours and so this starts decaying so the signal would diminish yes and also the conjugate starts getting eliminated from the mouse so this was basically the first project and that had completed in here and so we see that the TRC105 conjugation to graphene oxide it increases the tumor uptake also PET has a lot of good quantitative and non-invasive measurement of graphene oxide conjugates in the living mice and if we can further develop it we even need to perform some more experiments like some blocking experiments to confirm that graphene oxide and TRC105 conjugates are actually being directed to the tumor because of CD105 because of specificity for CD105 and not because the tumor that's the nature is just leaky and so antibodies tend to like you know get easy get into it easily and so if this thing can be developed further on so in future we might expect some image-rided drug delivery also from these conjugates or similar conjugates now my second project was development of antibody-based dual modality PET optical imaging agent in which we used an antibody the same antibody TRC105 the main reason why we want to do dual modality imaging is because as of now we do not have any modality a single imaging modality which is perfect in itself and would be enough to answer all your questions about diagnosis of a particular disease for example optical imaging it is it has good spatial resolution but it is very difficult to accurately quantify the fluorescence in our signals that we get and also it has two additional penetration with PET which is very sensitive and can be very easily quantified the problem is that it has poor spatial resolution the images are not very well resolved so if we can combine both of these we can get complimentary