The present invention relates to the field of anastasis, i.e., the process of reversal of apoptosis. More specifically, the present invention provides methods and compositions useful for studying anastasis. In one embodiment, the present invention provides an in vivo biosensor comprising (a) a transcription factor complex comprising the Gal4 transcription factor linked to an enzyme cleavable linker, wherein the transcription factor complex is tethered to the plasma membrane via a transmembrane domain; and (b) a reporter system comprising (1) a first nucleic acid encoding flippase operably linked to the upstream activating sequence that binds Gal4; and (2) a second nucleic acid comprising an FRT-flanked stop codon cassette separating a constitutive promoter and a fluorescent protein open reading frame.

A method for treating prostate cancer in a subject involves selecting a subject having prostate cancer and cytochrome c-deficiency, and administering, to the selected subject, a therapeutically effective amount of one or more agents capable of restoring cytochrome-c activity. Also presented is a method of inducing apoptosis in drug resistant cancer cells involving selecting drug resistant cancer cells having cytochrome-c deficiency, and administering to the selected cells, one or more agents that restore cytochrome-c activity in an amount effective to sensitize said cancer cells to drug induced apoptosis. A combination therapeutic comprising one or more agents increases cytochrome-c activity and efficacy of a chemotherapeutic agent. Another method involves selecting a subject having cancer, and obtaining a cell sample including tumor tissues/biopsy and blood samples from said subject, and further involves measuring cytochrome-c expression levels and Drp1 phosphorylation levels in said sample.

Disclosed herein are antibodies having binding specificity to the amino acid sequences Ala Ser Ser Gly Leu Thr Val Glu Val Asp (SEQ ID NO:1) and Thr Val Glu Val Asp (SEQ ID NO:14), and methods of detecting cell death in a sample, comprising contacting the sample with a first antibody specific for a C-terminal amino acid sequence Ala Ser Ser Gly Leu Thr Val Glu Val Asp (SEQ ID NO:1) or Thr Val Glu Val Asp (SEQ ID NO:14) of a CK18 protein fragment having a C-terminal amino acid sequence of Val Glu Val Asp (SEQ ID NO:2) and a second antibody that specifically binds an epitope that is present in both full-length CK18 and the CK18 protein fragment, and that does not overlap with SEQ ID NO:1 or SEQ ID NO:14, under conditions such that the CK18 protein fragment present in the sample specifically binds to the first antibody and the second antibody, wherein one of the antibodies is bound to a solid support and the other antibody is bound to a detection moiety capable of producing a signal; optionally removing any unbound or excess material; and detecting the signal from the detection moiety, wherein the signal is positively correlated with the presence of the CK18 protein fragment in the sample.

Immunoassays to detect the presence of Bcl-2 family heterodimeric complexes are described. The immunoassays are designed to detect one or more of BIM-Bcl-2, BIM-Bcl-xL, BIM-Mcl-1 and BAX-BAK heterodimers. The assays can be used, for example, to select a BH3 mimetic, or other drug targeting the apoptosis pathway, that is effective for treating a specific cancer, or to select a subject who is likely to respond to a particular BH3 mimetic.

A microfluidic device and method of assaying for immune cell exhaustion therewith are provided. The microfluidic device includes a moveable rod positioned across a chamber of a microfluidic device adjacent a first end thereof. Target cells are mixed into a hydrogel and the hydrogel is injected into the chamber about the moveable rod. The hydrogel is polymerized in. the chamber and the moveable rod is removed from the hydrogel so as to form a passageway in the hydrogel. The passageway is filled with a solution including immune cells. The immune cells migrate/diffuse into the hydrogel. A gradient of nutrients is formed in the chamber from. the first end to a second end of the chamber. One or more biopsies of the hydrogel may be taken at user selected location(s) of the chamber.

The disclosure provides methods of treating a tumor/cancer by administering naltrexone or an analogue thereof, followed by a recovery phase, and then administering a small molecule signaling inhibitor such as PI3-kinase inhibitors, AKT inhibitors, taxanes, antimetabolites, alkylating agents and/or cell cycle inhibitors. The disclosure also provides diagnostic methods for assessing a therapeutic response to the methods of treatment.

The present invention relates to the field of anastasis, i.e., the process of reversal of apoptosis. More specifically, the present invention provides methods and compositions useful for studying anastasis. The present invention also provides a biosensor comprising (a) a split transcription factor complex comprising one half of a split transcription factor linked to a transmembrane domain via an enzyme cleavable linker; (b) a split transcription factor comprising the other half of the split transcription factor linked to a MTS via an enzyme-cleavable linker; and (c) a reporter system comprising (1) a first nucleic acid encoding a site specific recombinase operably linked to the site specific sequence for the transcription factor; and (2) a second nucleic acid comprising a stop codon cassette flanked by site specific recombination sequences, wherein the split transcription factor is Gal 4 or split Q. In other embodiments, the recombinase is Cre or FLP.

Described herein are functional cell assays and methods for selecting a personalized anti-cancer treatment regimen that can improve treatment of cancer in a subject, identify resistance of the subject's cancer to one or more anti-cancer agents and/or validate the current drug treatment strategy.

Provided herein are compounds having the following structure (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.a, R.sup.b, and R.sup.c are as defined herein. Also provided are pharmaceutical compositions comprising compounds of structure (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and methods for use of compounds of structure (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, for treating diseases associated with overexpression of a cyclin-dependent kinase (CDK).

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The disclosure provides methods of treating a tumor/cancer by administering naltrexone or an analogue thereof, followed by a recovery phase, and then administering a small molecule signaling inhibitor such as PI3-kinase inhibitors, AKT inhibitors, taxanes, antimetabolites, alkylating agents and/or cell cycle inhibitors. The disclosure also provides diagnostic methods for assessing a therapeutic response to the methods of treatment.