Trojan circulating tumor cells
Provided herein are compositions and methods for engineering treatment resistant circulating tumor cells to generate immune stimulating or anti-cancer exosomes, which can be infused into a patient to self-home to the primary tumor and metastatic lesions. Also provided are compositions and methods for forming a population of engineered exosome-loaded dendritic cells, which can be infused into the patient to activate a systemic immune response. The compositions and methods allow for infiltration into solid tumors using “trojan” cells that are naturally adapted and mechanistically engineered to evade patients' defenses.
1 . A method of treating a cancer patient comprising:
(a) extracting circulating tumor cells from the cancer patient to produce a population of isolated circulating tumor cells;
(b) isolating a population of treatment resistant circulating tumor cells from the population of isolated circulating tumor cells;
(c) genetically engineering the treatment resistant circulating tumor cells to generate immune stimulating or anti-cancer exosomes to produce a population of genetically engineered treatment resistant circulating tumor cells;
(d) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient.
2 . The method of claim 1 , further comprising, either sequentially or simultaneously,
(a) harvesting exosomes from the population of treatment resistant circulating tumor cells to produce a population of isolated treatment resistant circulating tumor cell exosomes;
(b) incubating the population of isolated treatment resistant circulating tumor cell exosomes with dendritic cells to form a population of exosome-loaded dendritic cells; and
(c) infusing the population of exosome-loaded dendritic cells into the cancer patient.
3 . The method of claim 1 , wherein the population of isolated circulating tumor cells is extracted from a biological fluid, a fragmented tumor, a tumor suspension, a tissue suspension, a cell culture, an established cell line, or combinations thereof.
4 . The method of claim 1 , wherein extracting circulating tumor cells further comprises using a biological fluid filtration system, immunomagnetic separation, a system for identification and enumeration of circulating tumor cells from blood, immunoaffinity purification using antibodies, capture using aptamers, nanostructured surfaces for capture, size-based filtration, microfluidic separation, dielectrophoresis-based separation, or combinations thereof.
5 . The method of claim 1 , wherein the population of isolated circulating tumor cells is greater than about 1,000 cells.
6 . The method of claim 1 , wherein isolating the population of treatment resistant circulating tumor cells from the population of isolated circulating tumor cells further comprises:
exposing the population of isolated circulating tumor cells to anti-cancer agents for 1, or more days; and
measuring cell viability and or cell survival.
7 . The method of claim 1 , wherein the population of treatment resistant circulating tumor cells is isolated from the population of circulating tumor cells when one or more anti-cancer agents inhibit tumor cell growth, decrease cell viability, and/or decrease cell survival by about 50% or more when compared to a tumor cell whose growth, viability, and/or cell survival is inhibited or destroyed by one or more anti-cancer agents.
8 . The method of claim 1 , wherein the population of treatment resistant circulating tumor cells comprises circulating tumor cells that are chemo-resistive, targeted therapy resistive, immunotherapy resistive, or cells resistant to combinations of treatments.
9 . A method of treating a cancer patient comprising:
(a) extracting circulating tumor cells from a cancer patient to produce a population of isolated circulating tumor cells;
(b) isolating a population of treatment resistant circulating tumor cells from the population of isolated circulating tumor cells;
(c) harvesting exosomes from the population of isolated treatment resistant circulating tumor cells to produce a population of isolated treatment resistant circulating tumor cell exosomes;
(d) incubating the population of isolated treatment resistant circulating tumor cell exosomes with dendritic cells to form a population of exosome-loaded dendritic cells; and
(e) infusing the population of exosome-loaded dendritic cells into a cancer patient in need thereof.
10 . The method of claim 9 , further comprising:
(f) genetically engineering the treatment resistant circulating tumor cells to generate immune stimulating or anti-cancer exosomes to produce a population of genetically engineered treatment resistant circulating tumor cells; and
(g) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient in need thereof.
11 . The method of claim 10 , wherein (g) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient occurs simultaneously with (e) administering the population of exosome-loaded dendritic cells to the cancer patient in need thereof.
12 . The method of claim 10 , wherein (g) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient occurs prior to (e) infusing the population of exosome-loaded dendritic cells into the cancer patient in need thereof.
13 . The method of claim 10 , wherein (g) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient occurs after to (e) infusing the population of exosome-loaded dendritic cells into the cancer patient in need thereof.
14 . The method of claim 10 , wherein (g) administering the genetically engineered treatment resistant circulating tumor cells to the cancer patient and (e) infusing the population of exosome-loaded dendritic cells into the cancer patient in need thereof are separated by 2 or more hours.
15 . The method of claim 10 , wherein steps (f)-(g) are repeated 2 or more times.
16 . The method of claim 9 , wherein the population of isolated circulating tumor cells is greater than about 1,000 cells.
17 . The method of claim 9 , wherein the cancer patient and the cancer patient in need thereof are different patients.
18 . The method of claim 9 , wherein the cancer patient and the cancer patient in need thereof are the same patient.
19 . The method of claim 9 , wherein steps (a)-(e) are repeated 2 or more times.
20 . The method of claim 9 , wherein administering the population of exosome-loaded dendritic cells produces a localized response in the cancer patient in need thereof.