Phosphocofilin: cofilin co-localization intensity as a predictor of metastatic recurrence
Methods and products are provided for determining if a subject having a tumor is at risk of metastasis of the tumor. Specifically, the methods comprise detecting phosphorylated cofilin, and both phosphorylated and non-phosphorylated cofilin; quantifying the phosphorylated cofilin, and the total of phosphorylated and nonphosphorylated cofilin; and determining if a subject having the tumor is likely to experience metastasis of the tumor, based on the ratio of the amount of detected phosphorylated cofilin:total amount of phosphorylated and non-phosphorylated cofilin detected. Further disclosed are the types of tumor metastases that can be determined using the methods provided.
1. A method of treating a subject having a tumor comprising:
(A) identifying or having identified a subject as in need of an anti-metastatic treatment of a tumor, wherein the subject is identified as such by determining if an amount of phosphorylated cofilin bound: total amount of phosphorylated cofilin and non-phosphorylated cofilin bound colocalized in a sample of the tumor is in excess of a predetermined ratio value and indicative of metastasis, or a colocalization intensity value (IF) of an image of a sample obtained on the fluorescence imaging device within a colocalized area in the tumor is in excess of a predetermined IF value and indicative of metastasis, where a sample of the tumor has been contacted with (a) a fluorescent moiety-labeled agent that binds phosphorylated cofilin and (b) a fluorescent moiety-labeled agent which binds both phosphorylated cofilin and non-phosphorylated cofilin, and the method quantifies
(i) the phosphorylated cofilin bound by (a) and
(ii) the total of phosphorylated cofilin and non-phosphorylated cofilin bound by (b), and wherein the IF is the product of the intensity of fluorescence of the fluorescent moiety-labeled agent that binds phosphorylated cofilin and the fluorescence of the fluorescent moiety-labeled agent that binds both phosphorylated cofilin and non-phosphorylated cofilin at each pixel of the colocalized area,
wherein the fluorescent moiety-labeled agent that binds phosphorylated cofilin comprises an antibody, or antigen-binding fragment thereof, that specifically binds phosphorylated cofilin, and wherein the fluorescent moiety-labeled agent which binds to phosphorylated cofilin and to non-phosphorylated cofilin comprises an antibody, or antigen-binding fragment thereof, which binds phosphorylated cofilin and which also binds non-phosphorylated cofilin, and
(B) administering, to a subject so-identified in (A), an anti-metastatic therapy;
wherein the tumor is a breast tumor.
2. The method of claim 1 , wherein IF is intensity of (a) in pixels of an image of the sample multiplied by intensity of (b) in pixels of the image of the sample, optionally first masking out pixels with low Pearson distribution.
3. The method of claim 1 , wherein the anti-metastatic therapy comprises trastuzumab and/or bevacizumab.
4. The method of claim 1 , wherein the sample is a biopsy sample.
5. The method of claim 1 , wherein the sample has been surgically removed from a subject.
6. The method of claim 1 , wherein both the detectable agents each comprise a fluorescent moiety and wherein the detectable agent that binds to phosphocofilin comprises a different fluorescent moiety from the fluorescent moiety of the detectable agent that binds to phosphorylated cofilin and to non-phosphorylated cofilin.
7. The method of claim 1 , wherein the phosphorylated cofilin and non-phosphorylated cofilin are human cofilin.
8. The method of claim 1 , wherein the cofilin comprises SEQ ID NO:1.
9. The method of claim 1 , wherein the sample is a paraffin-embedded biopsy sample.
10. The method of claim 1 , wherein the fluorescent moiety-labeled agent which binds to phosphorylated cofilin and wherein the fluorescent moiety-labeled agent which binds to non-phosphorylated cofilin comprises an anti-pan-cofilin antibody.
11. The method of claim 1 , wherein the fluorescent moiety-labeled agent which specifically binds to phosphorylated cofilin comprises an anti-phospho-Ser-3-cofilin antibody.
12. The method of claim 1 , wherein the predetermined IF value-is determined from one or more subjects with malignant tumors which have not metastasized.
13. The method of claim 1 , wherein the system performs the method for determining if the colocalization intensity value (IF) of an image of a sample obtained on the fluorescence imaging device within a colocalized area in the tumor is in excess of a predetermined IF value and indicative of metastasis.