Early Cancer Detection And Enhanced Immunotherapy

Abstract

A method of therapy for a tumor or other pathology by administering a combination of thermotherapy and immunotherapy optionally combined with gene delivery. The combination therapy beneficially treats the tumor and prevents tumor recurrence, either locally or at a different site, by boosting the patient's immune response both at the time or original therapy and/or for later therapy. With respect to gene delivery, the inventive method may be used in cancer therapy, but is not limited to such use; it will be appreciated that the inventive method may be used for gene delivery in general. The controlled and precise application of thermal energy enhances gene transfer to any cell, whether the cell is a neoplastic cell, a pre-neoplastic cell, or a normal cell.

Claims

1. A cancer therapeutic method comprising administering to a patient having an early stage tumor a combination of thermotherapy and immunotherapy combined with gene therapy, the patient having a non-surgically accessible early stage tumor, where thermotherapy comprises systemically administering a tumor-antibody-coated nanoparticle, optionally also containing an agent that facilitates cell penetration and optionally coated with a thermosensitive polymer; and immunotherapy comprises systemically administering the patient's natural killer (NK) cells/dendritic cells pre-sensitized in vitro to the tumor.

2. The method of claim 1 where immunotherapy is administered at intervals to the patient after the initial therapy acting as a booster to the original immunotherapy and reduce or prevent tumor recurrences at a same or different site.

3. The method of claim 1 where immunotherapy further comprises administering NK cells/dendritic cells containing viral like particles (VLP).

4. The method of claim 1 where immunotherapy further comprises obtaining NK cells/dendritic cells grown in culture under light pulses with a tumor biomarker from blood or a tumor biopsy specimen containing tumor lysate, killed circulating tumor cells (ct cell), and tumor extracellular vesicles (ECV).

5. The method of claim 1 where thermotherapy includes exposing the tumor-antibody-coated nanoparticle to a light pulse at a frequency in the range of 20 Hz-60 Hz to decrease proliferation of the tumor cell.

6. The method of claim 3 where thermotherapy includes using a thermoacoustic unit to control a thermal energy delivery unit using a processor to maintain the nanoparticles at a predetermined temperature as a closed circuit once the nanoparticles have attached to the tumor cells, then controllably increasing the temperature to which the nanoparticles are exposed from 37 C. to 41 C.-43 C. for a predetermined desired time period resulting in melting a thermosensitive polymer coating the nanoparticles, releasing under control a conjugated VLP, medication/gene which are attached to the thermosensitive antibody coated nanoparticles locally at the desired site.

7. The method of claim 3 where immunotherapy comprises stimulating the patient's cellular immune response at the tumor site by (a) the released VLP providing tumor-localized immunotherapy, and (b) the released antigenic material from the thermotherapy-damaged tumor cells providing localized and non-localized immunotherapy functioning as an internal vaccination method.

8. The method of claim 7 where (b) additionally provides immune memory.

9. The method of claim 7 resulting in a temperature increase at any localized or non-localized site, the temperature increase permitting tumor imaging at the localized or non-localized site by photoacoustic technology.

10. The method of claim 1 where the anti-tumor antibody is specific for at least one tumor biomarker in the patient's blood.

11. The method of claim 1 where the antibodies from a patient's blood are to a tumor biomarker in the patient's blood.

12. The method of claim 6 where the polymer contains an inhibitory gene(s) and a CRISPR/cas9 complex to stimulate or modify the tumor genes upon release from the polymer at a desired temperature that is obtained by incremental increase from 37 C. to 43 C. permitting precision nanoparticle assisted thermotherapy and imaging (NATTI) to release the gene(s) and optional medicament(s) and/or checkpoint inhibitor(s) from the nanoparticles.

13. The method of claim 1 further comprising conjugating the nanoparticles with a chimeric receptor on B cells and with T-cells cultured in vitro and expressing an antigen for the chimeric receptor to target abnormal B cells seen in leukemia.

14. The method of claim 1 where the antibody is against an amyloid plaque.

15. The method of claim 1 further comprising performing precision nanoparticle assisted thermotherapy and generating a photoacoustic image of the cells to which the nanoparticles bind at an early cellular stage having a size less than four millimeters not easily detectable by radiographic imaging.

16. A method of simultaneous localized thermotherapy and vaccination, the method comprising administering a plurality of nanoparticles to a patient in need thereof, the administered nanoparticles having a size less than 10 nm in diameter and coated with an antitumor antibody, the patient provided nanoparticle assisted thermotherapy, the unbound nanoparticles undergoing renal elimination from the body within a few hours of administration, and the tumor-bound nanoparticles remaining in the patient.

17. A therapeutic method to treat a pathology implicating a cellular protein(s), the method comprising administering to a patient having a cellular pathology a combination of thermotherapy and immunotherapy, optionally with gene delivery, where thermotherapy comprises systemically administering an a antibody-coated nanoparticle, optionally also containing an agent that facilitates cell penetration and optionally coated with a thermosensitive polymer; immunotherapy comprises systemically administering the patient's natural killer (NK) cells/dendritic cells pre-sensitized in vitro to the cellular protein(s); and gene delivery, if used, comprises use of a CRISPR/cas9 and/or CRISPRi to replace or modify at least one genetic component in the cell.

18. The method of claim 17 where thermotherapy includes in vitro exposing the antibody-coated nanoparticle to a light pulse at a frequency in the range of 1 Hz-20 Hz to increase proliferation of cell.

19. The method of claim 17 wherein the pathological cellular protein(s) is an amyloid plaque and the patient has Alzheimer's disease, and/or is a protein from a microorganism and the patient is sub-responsive to therapy for that microorganism.

20. The method of claim 17, further comprising performing plasmaphoresis on the patient post-therapy to purify a patient's blood from toxins and cellular components generated by the therapy.

21. The method of claim 17, further comprising applying a strong pulse of light energy to a tube containing the patient's blood cells post-therapy to achieve a temperature up to 60 C. to kill immune cells containing quantum dots; passing the pulsed blood cells through a dielectrophoresis system to characterize and remove dead or live T-cells, sensitized killer cells, and tumor cells; and re-infusing the dielectrophoresis treated blood in the patient while simultaneously administering immunosuppressive agents, thus reducing the likelihood of a severe post-therapy autoimmune response in the patient.

Description

EXAMPLE 1

[0115] T cells and dendritic cells are obtained from a patient's blood, and grown in culture along with a tumor or other antigen, plus nanoparticles coated with thermosensitive polymers conjugated with antigen and VLP using culture methods known in the art.

[0116] The nanoparticle complex is injecting them along with checkpoint inhibitors and IL-2. The inventive method is applied, killing tumor cells, and increasing the response of T-cells and dendritic cell.

[0117] The patient's blood is assessed for new biomarkers from the dead cells.

[0118] The cultured T-cells and dendritic cells are harvested, along with the nanoparticle-coated antigen plus VLP or RNA or DNA phages. These are stored under appropriate conditions, and reinjected into the patient with low dose coated nanoparticles or systemic medicaments to be administered as needed, e.g., semi-annually, annually, biannually, etc. with repetition as needed. This is followed up with administering free circulating DNA and exosomes of circulating cells.

EXAMPLE 2

[0119] A checkpoint inhibitor is added to a thermosensitive polymer coating a nanoparticle for controlled release of the checkpoint inhibitor using the inventive NATTI to treat a patient with breast, colorectal, pancreatic, and/or ovarian cancer. A checkpoint inhibitor such as nivolumab is combined with nanoparticle assisted targeted immunotherapy for adaptive T-cell transfer to overcome the limitations of standard immunotherapy.

EXAMPLE 3

[0120] Nanoparticles are conjugated with a chimeric receptor, a CD19 protein that is found only on B cells, along with the T-cells cultured in vitro that expresses a chimeric antigen receptor (chimeric antigen receptor T (CAR T)-cells) to target abnormal B cells seen in leukemia. The reappearance of new biomarkers as neoantigens in these patients can be also treated in the postoperative period using the inventive method.

[0121] Plasmaphoresis is simultaneously performed or performed after treatment.

[0122] This example treats acute and chronic hematologic malignancies such as acute lymphoblastic leukemia, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, etc.

EXAMPLE 4

[0123] Nanoparticles are conjugated with all-trans retinoic acid (ATRA) and arsenic trioxide to target leukaemia cells in acute promyelocytic leukemia and used in the inventive method. The all-trans retinoic acid is released at the site of the tumor without exposing the entire body to the toxic medication, simultaneously, plasmophoresis is performed to clear all toxin released in the blood, along with leukemic cells. It is appreciated that other blood cell cancers are removed in the same session.

EXAMPLE 5

[0124] In a patient with a hematologic malignancy that is resistant to chemotherapeutic agents or immune therapy, NATTI is performed with gene delivery, along with chemotherapeutic agents, to target all immune cells initially without subjecting the patient to systemic heavy chemotherapy, followed by bone marrow transplantation, without exposing the entire body to systemic chemotherapy.

EXAMPLE 6

[0125] Nanoparticles are conjugated with RNA that contains an aptamer, ribosomes, and siRNA in a thermosensitive polymer and administered to using NATTI to target specific tumor cells.

EXAMPLE 7

[0126] The microenvironment of the cancer cell is modified by delivering medicaments that block the uptake of exosomal signals and prevent the uptake of ECV. Such medications include choloroquine, heparin, cytochalasin D, and ethyl-isopropyl amiloride. These medications are approved for patient use. The medicaments are provided using NATTI in conjunction with chemotherapeutic agents.

EXAMPLE 8

[0127] The inventive method provides nanoparticle assisted localized immunothermotherapy and thermotherapy for delivery of customized vaccines to target core mutations in a patient. The immune cells or T-cells that can attack those core mutations are identified via a cancer biomarker. The immune cells or T-cells are then cultured with the nanoparticles coated with thermosensitive particles and VLP and IL-2. The nanoparticles are injected into the patient, controllably heated using a thermal energy source, and imaged, for specific patient therapy.

EXAMPLE 9

[0128] Nanoparticles are conjugated with RNA phage VLP, which is generally stable up to about 50 C. VLPs of the related RNA phage PP7 are crosslinked with inter-subunit disulfide bonds, rendering them significantly more stable. They exhibit high immunogenicity. Such nanoparticles complement the inventive NATTI technology and can be employed in anti-cancer and antibacterial treatment. Lytic phages attach to receptors on the bacterial surface, inject their genetic material through the bacterial membrane, and overtake the bacterium's transcription and translation machinery to synthesize new phages.

EXAMPLE 10

[0129] To prevent a severe autoimmune response after tumor immunotherapy, one uses after or before extracorporeal plasmapheresis, the nanoparticle assisted thermotherapy and imaging system to apply heavy thermal energy to a tube containing blood cells and to achieve a temperature as high as 60 C. to kill the sensitized immune cells containing nanoparticles. Blood is then passed through a dielectrophoresis system to characterize and remove dead or live T-cells, sensitized killer cells, and tumor cells prior to re-infusing blood in the patient while simultaneously administering immunosuppressive agents, including biologics. This reduces the severe autoimmune response often seen after tumor immunotherapy.

[0130] The embodiments shown and described in the specification are only specific embodiments of the inventor who is skilled in the art and are not limiting in any way. Therefore, various changes, modifications, or alterations to those embodiments may be made without departing from the spirit of the invention in the scope of the following claims. The references cited are expressly incorporated by reference herein in their entirety.