1) Specific Aims
Introduction to your problem (breast cancer and cell cycle, make the audience think that
what you want to study is important). However, very little is known about … (what new information
will your study add to the current knowledge).
Therefore, I propose to … The current proposal will test the hypothesis that …
(be specific here) The hypothesis will be tested using … (brief sentence on methods and
The specific aims of this proposal are:
AIM 1: Specifically state the aim of your proposal in one sentence. Elaborate on this
aim, experiments proposed, what data you expect to collect and how that data will answer
AIM 2: Feel free to pose another aim if you like.
Briefly describe the significance of these outcomes to the field.
2) Background and Significance
Here you can go in depth about 1. Breast cancer 2. The cell cycle and cancer 3. The
significance of your channel in #2. You can include data and/or pictures from your
references to support your hypothesis.
For Example: Cardiotoxicity is a serious side effect
associated with a number of antiarrhythmic compounds11;28. More surprising is that a large
number of non-cardiac drugs are similarly associated with increased cardiac risk, despite
the fact that they were not developed with cardiac ion channels as the intended target. In
general, drug-induced cardiotoxicity is characterized by acute or subacute changes in the
electrocardiogram (ECG), including prolongation of the QT interval. Prolongation of the
QT interval is associated with an increased risk for torsade de pointe arrhythmia (TdP),
ventricular tachycardia (VT) or ventricular fibrillation (VF)20. Currently, acute ECG
changes have been associated with over 70 non-cardiac compounds, most of which
inhibit the cardiac potassium channel hERG/IKr thereby prolonging cardiac
Non-cardiac drugs, hERG block and QT prolongation
Block of hERG/IKr currents by a number of noncardiac drugs such as terfenadine and astemizole clearly established the link between druginduced LQTS and inherited LQTS caused by mutations in hERG, the gene responsible
for IKr potassium currents20;29. Mutations in hERG produce loss of function with a corresponding
reduction in repolarizing cardiac IKr potassium currents causing prolonged QT intervals. Acquired
LQTS is produced by non-cardiac drugs, and in nearly all cases is due to block
of hERG/IKr currents25. Similar to the situation in inherited LQTS, the reduction of IKr currents by
channel blockade in acquired LQTS prolongs the QT interval and is associated with an increased
risk for adverse
3) Research Design and Methods
Aim 1: Title of aim 1
For example: be specific about doses and/or concentrations of drugs and how you
will apply. Therefore, in the first series of experiments we will test the effects of a
range of concentrations of As2O3 and SSG on Ca2+ current in freshly isolated
Next be specific about how you will culture your cells and the conditions under
which you will record. For example: For these and other experiments involving overnight
exposure to test compounds, ventricular myocytes will be isolated and cultured overnight
in M199 medium with or without the test compound. Patch clamp recordings will be made
on the following day using solutions of the following composition (mM): NaCl
(137), CsCl (5.4), MgCl2 (1.8), CaCl2 (1.8), glucose (10), and HEPES (10), pH 7.4 for
extracellular; and CsMeSO4 (130), TEA Cl (20), MgCl2 (1), EGTA (10), HEPES
(10), MgATP (4), Tris-phosphocreatine (14), Tris-GTP (0.3), and 50 U/ml creatine
phosphokinase, pH 7.2 for intracellular. Currents will be elicited using 300 ms step
depolarizations from a holding potential of -40 mV (to inactivate Na+ and low-voltageactivated Ca2+ currents) over a range of -50 to +70 mV in 10 mV increments to record
current-voltage relations (I-V). To determine current densities, currents will be normalized
to membrane capacitance determined using the membrane test function
in pClamp software (Axon Instruments). Normalized I-Vs will be compared for
differences between control and treated cells in both magnitude and activation range.
Current traces will also be analyzed and compared for activation and inactivation time
constants. This analysis may also provide clues to the mechanism of action i.e., changes
in phosphorylation, channel number or gating properties. In addition, we will correlate
significant effects on Ca2+ current amplitudes with changes in APD (as shown in figure
2B) where appropriate.
Anticipated Potential Problems:
This should be self explanatory.
4) Literature Cited
This material may consist of step-by-step explanations on how to solve a problem or examples of proper writing, including the use of citations, references, bibliographies, and formatting. This material is made available for the sole purpose of studying and learning - misuse is strictly forbidden.
Ion Channel KCa3
KCa3 ion channel contributes to various processes in malignant progression and cancerogenesis (Mohr et al., 2019a). Consequently, the channel has increasingly become a centerstage in its involvement in several cancer entities, one of them being breast cancer. The potential of KCa3 as an anti-tumor target and its role in breast cancer in which the channel expresses itself based on the cell cycle (Mohr et al., 2019a). The most recent studies such as that by Steudel et al. (2017) demonstrate that the pharmacological or genetic inhibition of the KCa3 interferes with the progression of the breast tumor vitriol in an orthotopic breast cancer mouse model. Other studies...