Biomarkers for use in pancreatic cancer treatment

Abstract

Provided herein are methods of determining a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, and, based on the total number of mutations: (a) selecting a patient for treatment with a glycogen synthase kinase-3 inhibitor; (b) treating a patient with a glycogen synthase kinase-3 inhibitor; and / or (c) continuing to treat a subject with a glycogen synthase kinase-3 inhibitor.

Description

BIOMARKERS FOR USE IN PANCREATIC CANCER TREATMENTCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application No. 63 / 633,948, filed on April 15, 2024, the contents of which are hereby incorporated by reference in their entirety.FIELD

[0002] Provided herein are methods for selecting, treating, and monitoring a patient suffering from pancreatic cancer. In particular, the present disclosure provides methods of determining a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, and, based on the total number of mutations: (a) selecting a patient for treatment with a glycogen synthase kinase-3 inhibitor; (b) treating a patient with a glycogen synthase kinase-3 inhibitor; and / or (c) continuing to treat a subject with a glycogen synthase kinase-3 inhibitor.BACKGROUND

[0003] Metastatic pancreatic cancer poses a formidable challenge in the realm of oncology due to its aggressive nature and limited treatment options. Despite advancements in cancer research and treatment modalities, pancreatic cancer continues to rank among the deadliest malignancies globally. Its high mortality rate is primarily attributed to the lack of early symptoms and effective screening methods, leading to delayed diagnosis and subsequent metastasis to distant organs. Advanced metastatic pancreatic cancer, characterized by the spread of malignant cells beyond the pancreas to vital organs like the liver, lungs, or peritoneum, presents clinicians with a daunting clinical scenario. The metastatic spread of pancreatic cancer to distant sites profoundly impacts patient prognosis and treatment effectiveness. Metastases to critical organs often lead to functional impairments, severe complications, and a decline in overall health status. Moreover, the aggressive nature of metastatic pancreatic cancer contributes to its resistance to conventional treatment modalities, such as chemotherapy, targeted therapy, and immunotherapy. Despite efforts to develop novel therapeutic strategies, includingcombination therapies and personalized medicine approaches, the survival rates for patients with advanced metastatic pancreatic cancer remain dismal, with only a fraction of patients surviving beyond five years post-diagnosis.

[0004] In recent years, there has been growing interest in targeting specific molecular pathways implicated in pancreatic cancer pathogenesis to improve treatment efficacy and patient outcomes. One such pathway of interest is the glycogen synthase kinase-3 (GSK-3) signaling pathway. GSK-3 is a multifunctional protein kinase involved in various cellular processes, including cell proliferation, differentiation, apoptosis, and metabolism. Dysregulation of GSK-3 activity has been implicated in the development and progression of various cancers, including pancreatic cancer.

[0005] Preclinical studies have demonstrated that aberrant GSK-3 activity can promote tumor growth, invasion, and metastasis in pancreatic cancer cells. Consequently, GSK-3 has emerged as a promising therapeutic target for cancer treatment, including pancreatic cancer. Inhibitors of GSK-3 have shown efficacy in preclinical models of pancreatic cancer by inhibiting tumor cell proliferation, inducing apoptosis, and suppressing tumor angiogenesis and metastasis. Additionally, the inclusion of predictive cancer biomarkers in these methods is of paramount importance, as they can further enhance the efficacy of GSK-3 inhibitors by aiding in the prediction of therapy outcomes, assessment of tumor response to treatment, determination of prognosis, patient stratification for initial therapies, and optimization of treatment strategies tailored to individual patients.

[0006] However, despite promising preclinical data, the clinical translation of GSK-3 inhibitors as anticancer agents, particularly in the context of advanced metastatic pancreatic cancer, remains a subject of ongoing research. Likewise, the lack of predictive biomarkers for patient selection and response assessment poses a significant challenge in the successful implementation of cancer treatments in clinical practice. Without the use of predictive biomarkers, the efficacy of cancer treatment may be compromised, leading to suboptimal outcomes for patients. While there is a rationale for targeting GSK-3 in pancreatic cancer therapy, the clinical efficacy and safety of GSK-3 inhibitors, either as monotherapy or in combination with standard treatments and the use of predictive cancer biomarkers, need to be further evaluated.SUMMARY

[0007] Embodiments of the present disclosure include methods of determining a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, and, based on the total number of mutations: (a) selecting a patient for treatment with a glycogen synthase kinase-3 inhibitor; (b) treating a patient with a glycogen synthase kinase-3 inhibitor; and / or (c) continuing to treat a subject with a glycogen synthase kinase-3 inhibitor.

[0008] Provided herein are methods of treating a subject suffering from pancreatic cancer, the method comprising the steps of: receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, wherein the at least one gene is KRAS, TP53, CDKN2A, or a combination thereof; and administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0; or not administering the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

[0009] In some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least two genes.

[0010] hi some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

[0011] In certain embodiments, the method comprises: administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinasc-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes;administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

[0012] In some embodiments, the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

[0013] In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, or any combinations thereof.

[0014] In some embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0015] In certain embodiments, the subject is treated with a combination of a glycogen synthase kinase-3beta inhibitor and a chemotherapeutic agent.

[0016] In further embodiments, the chemotherapeutic agent is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan / liposomal irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan / liposomal irinotecan, leucovorin and fluorouracil.

[0017] In certain embodiments, the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

[0018] In some embodiments, the glycogen synthase kinase-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

[0019] In further embodiments, wherein the glycogen synthase kinase-3 inhibitor is elraglusib.

[0020] In some embodiments, the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

[0021] In certain embodiments, the total number of mutations in the at least one gene is determined using a sequencing assay.

[0022] Provided herein are methods of selecting a subject suffering from pancreatic cancer for treatment with a glycogen synthase kinase-3 inhibitor, the method comprising the steps of: receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, wherein the at least one gene is KRAS, TP53, CDKN2A or a combination thereof; and selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

[0023] In some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least two genes.

[0024] In some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

[0025] In certain embodiments, the method comprises: selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; selecting a subject for treatment with a glycogen synthase kinasc-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; selecting a subject for treatmentwith a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

[0026] In some embodiments, the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

[0027] In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a scrum sample, a urine sample, or any combinations thereof.

[0028] In some embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0029] In certain embodiments, the subject is treated with a combination of a glycogen synthase kinase-3 inhibitor and a chemotherapeutic agent.

[0030] In further embodiments, the chemotherapeutic agent isa a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan, leucovorin and fluorouracil.

[0031] In certain embodiments, the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

[0032] In some embodiments, the glycogen synthase kinase-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

[0033] In further embodiments, the glycogen synthase kinase-3 inhibitor is elraglusib.

[0034] In some embodiments, the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

[0035] In certain embodiments, the total number of mutations in the at least one gene is determined using a sequencing assay.

[0036] Provided herein are methods of monitoring a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor, the method comprising the steps of: receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor, wherein the at least one gene is KRAS, TP53, CDKN2A or a combination thereof; and continuing to treat the subject with the glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0; or discontinuing treatment with the glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

[0037] hr some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least one gene.

[0038] In some embodiments, the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

[0039] In certain embodiments, the method comprises: continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or discontinuing treatment with a glycogensynthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

[0040] In some embodiments, the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

[0041] In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, or any combinations thereof.

[0042] In some embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0043] In certain embodiments, the subject is treated with a combination of a glycogen synthase kinase-3 inhibitor and a chemotherapeutic agent.

[0044] In further embodiments, the chemotherapeutic agent is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan, leucovorin and fluorouracil.

[0045] In certain embodiments, wherein the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

[0046] In some embodiments, the glycogen synthase kinasc-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

[0047] In further embodiments, the glycogen synthase kinase-3 inhibitor is elraglusib.

[0048] In certain embodiments, the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

[0049] In some embodiments, the total number of mutations in the at least one gene is determined using a sequencing assay.BRIEF DESCRIPTION OF THE DRAWINGS

[0050] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0051] Having thus described the presently disclosed subject matter in general terms, reference will now be made to the accompanying Figures, which are not necessarily drawn to scale, and wherein:

[0052] FIG. 1A-B: KRAS-mut in GnP and GnP+Elra-treated pancreatic cancer patients.(A) shows the overall survival (OS) in KRAS-wild type (wt) vs KRAS-mutation (mut) GnP- treated patients and (B) OS in KRAS-wt vs KRAS-mut GnP+Elra (lx / w+2x / w)-treated patients.

[0053] FIG. 2A-B: TP53-mut in GnP and GnP+Elra-treated pancreatic cancer patients.(A) shows the overall survival (OS) in TP53-wild type (wt) vs TP53- mutation (mut) GnP-treated patients and (B) OS in TP53-wt vs TP53- mut GnP+Elra (lx / w+2x / w)-treated patients.

[0054] FIG. 3A-B: CDKN2A-mut in GnP and GnP+Elra-treated pancreatic cancer. (A) shows the overall survival (OS) in CDKN2A-wild type (wt) vs CDKN2A- mutation (mut) GnP- treated patients and (B) OS in CDKN2A-wt vs CDKN2A- mut GnP+Elra (lx / w+2x / w)-treated patients.

[0055] FIG. 4A-B: Double-mut (KRAS-mut + TP53-mut) in GnP and GnP+Elra-treated pancreatic cancer patients. (A) shows the overall survival (OS) in Double-mutation (mut) vs non-double-mut GnP-treated patients and (B) OS in Double mutation (mut) vs non-double-mut GnP+Elra (lx / w+2x / w)-treated patients.

[0056] FIG. 5A-B: Triple-mut (KRAS, TP53, CDKN2A) in GnP and GnP+Elra-treated pancreatic cancer patients. (A) shows the overall survival (OS) in Triple- mutation (mut) vs non-triple-mut GnP-treated patients and (B) OS in Triple-pos vs non-triple-pos GnP+Elra (lx / w+2x / w)-treated patients.DETAILED DESCRIPTION

[0057] The present disclosure provides methods of determining a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, and, based on the total number of mutations: (a) selecting a patient for treatment with a glycogen synthase kinase-3 inhibitor; (b) treating a patient with a glycogen synthase kinase-3 inhibitor; and / or (c) continuing to treat a subject with a glycogen synthase kinase-3 inhibitor. The at least one gene used in the methods described herein is KRAS, TP53, CDKN2A, or a combination thereof.DEFINITIONS

[0058] The terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms "a," "and," and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments "comprising," "consisting of," and "consisting essentially of," the embodiments or elements presented herein, whether explicitly set forth or not.

[0059] For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 69, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated. Unless otherwise defined herein, scientific, and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear; in the event, however of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0060] As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, e.g., the inclusion of at least one, butalso including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of” or “exactly one of’ or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (e.g., “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of’ “only one of’ or “exactly one of.”

[0061] As used herein, the term "administering" means either directly administering a compound or composition of the present invention, or administering a prodrug, derivative or analog which will form an equivalent amount of the active compound or substance within the body.

[0062] As used herein, the term “agent” is used herein to denote a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues that are suspected of having therapeutic properties. The agent may be purified, substantially purified, or partially purified.

[0063] As used herein, the term “combination” is used in its broadest sense and means that a subject is administered at least two agents, more particularly a glycogen synthase kinase-3 inhibitor and at least one other therapeutic agent. More particularly, the term “in combination” refers to the concomitant administration of two (or more) active agents for the treatment of a, e.g., single disease state. As used herein, the active agents may be combined and administered in a single dosage form, may be administered as separate dosage forms at the same time, or may be administered as separate dosage forms that are administered alternately or sequentially on the same or separate days. In one embodiment of the presently disclosed subject matter, the active agents arc combined and administered in a single dosage form. In another embodiment, the active agents are administered in separate dosage forms (e.g., wherein it is desirable to vary the amount of one but not the other). The single dosage form may include additional active agents for the treatment of the disease state.

[0064] As used herein, the term “elraglusib” or “9-1NG-41” as used interchangeably herein, refers to a Glycogen Synthase Kinase-3 (GSK-3) inhibitor having the chemical name 3-(5- Fluoro-benzofuran-3-yl)-4-(5-methyl-5H-[l,3]dioxolo[4,5f]indol-7-yl)-pyrrole-2, 5-dione and themolecular formula C22H13N2O5F. The chemical structure of elraglusib is shown below in Formula I.Formula I

[0065] As used herein, the term "Glycogen Synthase Kinase-3” or "GSK-3" are used interchangeably herein refers to a serine / threonine, proline-directed kinase involved in a diverse array of signaling pathways, including glycogen synthesis and cellular adhesion. GSK-3 has two isoforms (a and p) and is involved in glycogen metabolism. The term refers to proteins of the same or similar names and functional fragments and homologs thereof. The term also includes any recombinant or naturally occurring form of, or variants thereof that maintain GSK- 3 activity (e.g., within at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% activity compared to GSK-3.

[0066] As used herein, the term “inhibit,” or “inhibitor” and grammatical derivations thereof, refers to the ability of a glycogen synthase kinase-3 inhibitor, to block, partially block, interfere, decrease GSK-3, such as of GSK-3alpha, GSK-3beta, or GSK3-alpha and GSK-3beta. Thus, one of ordinary skill in the ail would appreciate that the term “inhibit” or “inhibitor” encompasses a complete and / or partial decrease in GSK-3 (such as, GSK-3alpha, GSK-3beta, or GSK3-alpha and GSK-3beta), e.g., a decrease by at least 10%, in some embodiments, a decrease by at least 20%, 30%, 50%, 75%, 95%, 98%, and up to and including 100%. Examples of GSK-3 inhibitors include elraglusib, Laduviglusib, SB216763, AT7519, TWS119, CHIR-98014, LY2090314, Tideglusib, SB415286, AR-A014418, 1-Azakenpaullone, IM-12, CP21R7, TDZD-8, AZD2858, Indirubin, AZD1080, Bikinin, Resibufogenin, 7-bromoindirubin-3-oxime, 6-bromoindirubin-3- oxime, MAZ51, PF-04802367, WAY-119064, BRD0705, or any combination thereof.

[0067] As used herein, the term “gene” refers to a nucleic acid (e.g., DNA or RNA) sequence that comprises coding sequences necessary for the production of an RNA, or of a polypeptide or its precursor. A functional polypeptide can be encoded by a full-length coding sequence or by any portion of the coding sequence as long as the desired activity or functional properties (e.g., enzymatic activity, ligand binding, signal transduction, etc.) of the polypeptide are retained.

[0068] As used herein, the terms “patient” or “subject” refer to organisms to be subject to various tests provided by the technology. The term “subject” includes animals, preferably mammals, including humans. In a preferred embodiment, the subject is a primate. In an even more preferred embodiment, the subject is a human. Further with respect to diagnostic methods, a preferred subject is a vertebrate subject. A preferred vertebrate is warm-blooded; a preferred warm-blooded vertebrate is a mammal. A preferred mammal is most preferably a human. As used herein, the term “subject1includes both human and animal subjects. Thus, veterinary therapeutic uses are provided herein. As such, the present technology provides for the diagnosis of mammals such as humans, as well as those mammals of importance due to being endangered, such as Siberian tigers; of economic importance, such as animals raised on farms for consumption by humans; and / or animals of social importance to humans, such as animals kept as pets or in zoos. Examples of such animals include but are not limited to: carnivores such as cats and dogs; swine, including pigs, hogs, and wild boars; ruminants and / or ungulates such as cattle, oxen, sheep, giraffes, deer, goats, bison, and camels; pinnipeds; and horses. Thus, also provided is the diagnosis and treatment of livestock, including, but not limited to, domesticated swine, ruminants, ungulates, horses (including race horses), and the like.

[0069] As used herein, whether by itself or in conjunction with another term or terms, it should be understood that the phrases "method of treating" and "method of treatment" may be used interchangeably with the phrase "for use in the treatment of a particular disease”.

[0070] As used herein, whether used alone or in conjunction with another term or terms, "therapeutic" and "therapeutically effective amount" refer to an amount of a compound, active agent, or composition that (a) treats a particular condition, symptom, disorder, or disease described herein; (b) attenuates, ameliorates, or eliminates one or more symptoms of a particular condition, disorder, or disease described herein; (c) delays the onset or relapse (reoccurrence) of a particular condition, symptom, disorder, or disease described herein; (d) prevents the onset of aparticular condition, symptom, disorder, or disease described herein. It should be understood that the terms "therapeutic" and "therapeutically effective" encompass any one of the aforementioned effects (a)-(d), either alone or in combination with any of the others (a)-(d).

[0071] As used herein, the term “sample” is used in its broadest sense. In one sense, it is meant to include a specimen obtained from any source, including biological samples. Biological samples may be obtained from animals (including humans) and encompass fluids, solids, tissues, and gases. Such examples are not however to be construed as limiting the sample types. As used herein, “sample” and “biological sample” are used interchangeably.

[0072] In some aspects, a sample is a fluid sample such as a liquid sample. Examples of liquid samples that may be assayed include bodily fluids (e.g., is a whole blood sample, a plasma sample, a serum sample, a urine sample, a bile sample, a pancreatic juice sample, a pleural fluid or ascites sample, a lymph sample, a peritoneal fluid sample, a saliva sample, a cerebrospinal fluid sample, a stool sample or any combination thereof), samples from water sources or sewage samples; and food samples.

[0073] Viscous liquid, semisolid, or solid specimens may be used to create liquid solutions, eluates, suspensions, or extracts that can be samples. For example, throat or genital swabs may be suspended in a liquid solution to make a sample. In other aspects, the biological sample is a tumor biopsy sample, such as, a brush cytology biopsy sample, a piece of tumor tissue or combinations thereof.

[0074] Samples can include a combination of liquids, solids (e.g., tissues), or any combination thereof (e.g., a suspension of lysed or unlysed cells in a buffer or solution). Samples can comprise biological materials, such as cells, microbes, organelles, and biochemical complexes. Liquid samples can be made from solid, semisolid, or highly viscous materials, such as fecal matter, tissues, organs, biological fluids, or other samples that arc not fluid in nature. In some aspects, the liquid sample is For example, solid or semisolid samples can be mixed with an appropriate solution, such as a buffer, a diluent, and / or extraction buffer. The sample can be macerated, frozen and thawed, or otherwise extracted to form a fluid sample. Residual particulates may be removed or reduced using conventional methods, such as filtration or centrifugation.

[0075] A variety of cell types, tissue, or bodily fluid may be utilized to obtain a sample or biological sample. Such cell types, tissues, and fluid may include sections of tissues such as biopsy and autopsy samples, oropharyngeal specimens, nasopharyngeal specimens, nasal mucus specimens, frozen sections taken for histologic purposes, blood (such as whole blood, dried blood spots, etc.), plasma, serum, red blood cells, platelets, an anal sample (such as an anal swab specimen), interstitial fluid, cerebrospinal fluid, etc. Cell types and tissues may also include lymph fluid, cerebrospinal fluid, or any fluid collected by aspiration. A tissue or cell type (such as a tumor biopsy sample) may be provided by removing a sample of cells from a human and a non-human animal, but can also be accomplished by using previously isolated cells (e.g., isolated by another person, at another time, and / or for another purpose). Archival tissues, such as those having treatment or outcome history, may also be used.

[0076] As used herein, whether by themselves or in conjunction with another term or terms, "treats," "treating," "treated," and "treatment," refer to and include ameliorative, palliative, and / or curative uses and results, or any combination thereof. In other embodiments, the methods described herein can be used prophylactically, that is, preventatively. It should be understood that "prophylaxis" or a prophylactic use or result do not refer to nor require absolute or total prevention (i.e., a 100% preventative or protective use or result). As used herein, prophylaxis or a prophylactic (preventative) use or result refers to uses and results in which administration of a compound or composition diminishes or reduces the severity of a particular condition, symptom, disorder, or disease described herein; diminishes or reduces the likelihood of experiencing a particular condition, symptom, disorder, or disease described herein; or delays the onset or relapse (reoccurrence) of a particular condition, symptom, disorder, or disease described herein; or any combination of the foregoing.

[0077] As used herein, the term “wild-type” when made in reference to a gene refers to a gene that has the characteristics of a gene isolated from a naturally occurring source. The term “wildtype” when made in reference to a gene product refers to a gene product that has the characteristics of a gene product isolated from a naturally occurring source. The term “wildtype” when made in reference to a protein refers to a protein that has the characteristics of a naturally occurring protein. The term “naturally-occurring” as applied to an object refers to the fact that an object can be found in nature. For example, a polypeptide or polynucleotide sequencethat is present in an organism (including viruses) that can be isolated from a source in nature, and which has not been intentionally modified by the hand of a person in the laboratory is naturally- occurring. A wild-type gene is often that gene or allele that is most frequently observed in a population and is thus arbitrarily designated the “normal” or “wild-type” form of the gene. In contrast, the term “modified” or “mutant” when made in reference to a gene or to a gene product refers, respectively, to a gene or to a gene product that displays modifications in sequence and / or functional properties (e.g., altered characteristics) when compared to the wild-type gene or gene product. It is noted that naturally-occurring mutants can be isolated; these are identified by the fact that they have altered characteristics when compared to the wild-type gene or gene product.Methods of Treating a Patient Suffering from Pancreatic Cancer

[0078] Provided herein are methods of treating a subject suffering from pancreatic cancer.The first step of the method involves receiving information on the total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer. The at least one gene is KRAS, TP53, CDKN2A, or any combination thereof. In some aspects, the total number of mutations is determined in at least two genes. In other aspects, the total number of mutations is determined in at least three genes.

[0079] In some embodiments, the biological sample obtained from a subject suffering from pancreatic cancer is a liquid sample, a tumor biopsy sample, a tissue biopsy sample, a bone marrow sample or a combination thereof. In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, a bile sample, a pancreatic juice sample, a pleural fluid or ascites sample, a lymph sample, a peritoneal fluid sample, a saliva sample, a cerebrospinal fluid sample, a stool sample or any combinations thereof. In yet other embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0080] Once a biological sample is obtained from a subject suffering from pancreatic cancer, the total number of mutations of at least one gene is determined using routine techniques known in the ail. In some aspects, the total number of mutations of at least one gene, at least two genes, and / or at least three genes can be determined using one or more sequencing assays. Examples of sequencing assays that can be used in the methods described herein include: whole genome sequencing (WGS), whole exome sequencing (WES), targeted sequencing, RNA sequencing(RNA-Seq), chromatin immunoprecipitation sequencing (ChlP-Seq), DNA methylation sequencing (Methyl-Seq), metagenomic sequencing, single-cell sequencing, or any combination thereof. In some aspects, the total number of mutations is determined for the KRAS gene. In other aspects, the total number of mutations is determined for the TP53 gene. In still other aspects, the total number of mutations is determined for the CDKN2A gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the TP53 gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the CDKN2A gene. In still other aspects, the total number of mutations is determined for the TP53 gene and the CDKN2A gene. In still yet other aspects, the total number of mutations is determined for the KRAS gene, the TP53 gene and the CDKN2A gene.

[0081] Once the total number of mutations in at least one KRAS, TP53, and / or CDLM2A gene is determined, the next step of the method involves making a decision of whether to administer a glycogen synthase kinase-3 inhibitor to the subject. Specifically, in some aspects, a glycogen synthase kinase-3 inhibitor is administered to the subject if the total number of mutations in at least one gene is zero. In other aspects, a glycogen synthase kinase-3 inhibitor is not administered to the subject if the total number of mutations in at least one gene is 1.

[0082] hi certain embodiments, the method comprises: administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinasc-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or not 1administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

[0083] In some embodiments, if pursuant to the above method, a determination is made that the subject is to be administered a glycogen synthase kinase-3 inhibitor, the glycogen synthase kinase-3 inhibitor administered to the subject can be a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

[0084] In still other embodiments, if pursuant to the above method, a determination is made that the subject is be administered a glycogen synthase kinase-3 inhibitor, the glycogen synthase kinase-3 inhibitor administered is elraglusib. In still other aspects, the subject is administered elraglusib in an amount of about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, about 21 mg / kg, about 22 mg / kg, about 23 mg / kg, about 24 mg / kg, about 25 mg / kg, about 26 mg / kg, about 27 mg / kg, about 28 mg / kg, about 29 mg / kg, about 30 mg / kg, about 31 mg / kg, about 32 mg / kg, about 33 mg / kg, about 34 mg / kg, about 35 mg / kg, about 36 mg / kg, about 37 mg / kg, about 38 mg / kg, about 39 mg / kg, about 40 mg / kg, about 41 mg / kg, about 42 mg / kg, about 43 mg / kg, about 44 mg / kg about 45 mg / kg, about 46 mg / kg, about 47 mg / kg, about 48 mg / kg, about 49 mg / kg or about 50 mg / kg, once, twice, three, four, or five times per week, or daily.

[0085] In some embodiments, the glycogen synthase kinase-3 inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthasekinase kinase-3alpha inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthase kinase kinase-3beta inhibitor is administered in combination with one or more chemotherapeutic agents. In still other aspects, elraglusib is administered in combination with one or more chemotherapeutic agents.

[0086] In certain embodiments, the chemotherapeutic agents used in the combination is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan / liposomal irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan / liposomal irinotecan, leucovorin and fluorouracil. In certain embodiments, the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

[0087] Methods of Selecting a Patient Suffering from Pancreatic Cancer

[0088] In other embodiments, provided herein are methods of selecting a patient suffering from pancreatic cancer for treatment with a glycogen synthase kinase-3 inhibitor. The first step of the method involves receiving information on the total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer. The at least one gene is KRAS, TP53, CDKN2A, or any combination thereof. In some aspects, the total number of mutations is determined in at least two genes. In other aspects, the total number of mutations is determined in at least three genes.

[0089] In some embodiments, the biological sample obtained from a subject suffering from pancreatic cancer is a liquid sample, a tumor biopsy sample, a tissue biopsy sample, a bone marrow sample or a combination thereof. In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, a bile sample, a pancreatic juice sample, a pleural fluid or ascites sample, a lymph sample, a peritoneal fluid sample, a saliva sample, a cerebrospinal fluid sample, a stool sample or any combinations thereof. In yet other embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0090] Once a biological sample is obtained from a subject suffering from pancreatic cancer, the total number of mutations of at least one gene is determined using routine techniques known in the art. In some aspects, the total number of mutations of at least one gene, at least two genes, and / or at least three genes can be determined using one or more sequencing assays. Examples of sequencing assays that can be used in the methods described herein include: whole genomesequencing (WGS), whole exome sequencing (WES), targeted sequencing, RNA sequencing (RNA-Seq), chromatin immunoprecipitation sequencing (ChlP-Seq), single-cell sequencing, or any combination thereof. In some aspects, the total number of mutations is determined for the KRAS gene. In other aspects, the total number of mutations is determined for the TP53 gene. In still other aspects, the total number of mutations is determined for the CDKN2A gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the TP53 gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the CDKN2A gene. In still other aspects, the total number of mutations is determined for the TP53 gene and the CDKN2A gene. In still yet other aspects, the total number of mutations is determined for the KRAS gene, the TP53 gene and the CDKN2A gene.

[0091] Once the total number of mutations in at least one KRAS, TP53, and / or CDLM2A gene is determined, the next step of the method involves making a decision of whether to select the subject for treatment with a glycogen synthase kinase-3 inhibitor. Specifically, in some aspects, a subject is selected for treatment with a glycogen synthase kinase-3 inhibitor if the total number of mutations in at least one gene is zero. In other aspects, a subject is not selected for treatment with a glycogen synthase kinase-3 if the total number of mutations in at least one gene is 1.

[0092] In certain embodiments, the method comprises: selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or not selecting a subject for treatment with aglycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

[0093] In some embodiments, if pursuant to the above method, a determination is made that the subject is to be selected for treatment with a glycogen synthase kinase-3 inhibitor, the subject is administered a therapeutically effective amount of a glycogen synthase kinase-3 inhibitor. In some aspects, the subject is administered a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

[0094] hi still other embodiments, if pursuant to the above method, a determination is made that the subject is be to be selected for treatment with a glycogen synthase kinase-3 inhibitor, the glycogen synthase kinase-3 inhibitor administered is elraglusib. In still other aspects, the subject is administered elraglusib in an amount of about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, about 21 mg / kg, about 22 mg / kg, about 23 mg / kg, about 24 mg / kg, about 25 mg / kg, about 26 mg / kg, about 27 mg / kg, about 28 mg / kg, about 29 mg / kg, about 30 mg / kg, about 31 mg / kg, about 32 mg / kg, about 33 mg / kg, about 34 mg / kg, about 35 mg / kg, about 36 mg / kg, about 37 mg / kg, about 38 mg / kg, about 39 mg / kg, about 40 mg / kg, about 41 mg / kg, about 42 mg / kg, about 43 mg / kg, about 44 mg / kg about 45 mg / kg, about 46 mg / kg, about 47 mg / kg, about 48 mg / kg, about 49 mg / kg or about 50 mg / kg, once, twice, three, four or five times per week, or daily.

[0095] In some embodiments, the glycogen synthase kinase-3 inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthase kinase kinase-3alpha inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthase kinase kinase-3beta inhibitor is administered in combination with one or more chemotherapeutic agents. In still other aspects, elraglusib is administered in combination with one or more chemotherapeutic agents.

[0096] In certain embodiments, the chemotherapeutic agents used in the combination is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan / liposomal irinotecan and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan / liposomal irinotecan, leucovorin and fluorouracil. In certain embodiments, the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

[0097] Methods of Monitoring a Patient Suffering from Cancer and Receiving Treatment with a Glycogen Synthase Kinase-3 Inhibitor

[0098] In other embodiments, also provided herein are methods of monitoring a patient suffering from cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor. The first step of the method involves receiving information on the total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor. The at least one gene is KRAS, TP53, CDKN2A, or any combination thereof. In some aspects, the total number of mutations is determined in at least two genes. In other aspects, the total number of mutations is determined in at least three genes.

[0099] hi some embodiments, the biological sample obtained from a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor is a liquid sample, a tumor biopsy sample, a tissue biopsy sample, a bone marrow sample or a combination thereof. In further embodiments, the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, a bile sample, a pancreatic juice sample, a pleural fluid or ascites sample, a lymph sample, a peritoneal fluid sample, a saliva sample, a cerebrospinal fluid sample, a stool sample or any combinations thereof. In yet other embodiments, the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

[0100] Once a biological sample is obtained from a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor, the total number of mutations of at least one gene is determined using routine techniques known in the art. In some aspects, the total number of mutations of at least one gene, at least two genes, and / or at least three genes can be determined using one or more sequencing assays. Examples of sequencing assays that can be used in the methods described herein include: whole genome sequencing (WGS), whole exome sequencing (WES), targeted sequencing, RNA sequencing (RNA-Seq), chromatin immunoprecipitation sequencing (ChlP-Seq), single-cell sequencing, or any combination thereof. In some aspects, the total number of mutations is determined for the KRAS gene. In other aspects, the total number of mutations is determined for the TP53 gene. In still other aspects, the total number of mutations is determined for the CDKN2A gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the TP53 gene. In still further aspects, the total number of mutations is determined for the KRAS gene and the CDKN2A gene. In still other aspects, the total number of mutations is determined for the TP53 gene and the CDKN2A gene. In still yet other aspects, the total number of mutations is determined for the KRAS gene, the TP53 gene and the CDKN2A gene.

[0101] Once the total number of mutations in at least one KRAS, TP53, and / or CDLM2A gene is determined, the next step of the method involves making a decision of whether to continue to treat the subject with a glycogen synthase kinase-3 inhibitor. Specifically, in some aspects, treatment with the glycogen synthase kinase-3 inhibitor is continued for the subject if the total number of mutations in at least one gene is zero. In other aspects, treatment with the glycogen synthase kinase-3 inhibitor is discontinued for the subject if the total number of mutations in at least one gene is 1.

[0102] In certain embodiments, the method comprises: continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; continuingto treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.[001031 In some embodiments, if pursuant to the above method, it is determined that the patient is to continue with treatment with a glycogen synthase kinase-3 inhibitor, then the patient can continue on their specific dosing regimen. For example, the subject can continue on their specific dosing regimen of elraglusib. Alternatively, the dosing regimen of the subject can be altered. For example, if the subject is being treated with elraglusib, the dosing regimen can be altered such that the subject is administered elraglusib in an amount of about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, about 21 mg / kg, about 22 mg / kg, about 23 mg / kg, about 24 mg / kg, about 25 mg / kg, about 26 mg / kg, about 27 mg / kg, about 28 mg / kg, about 29 mg / kg, about 30 mg / kg, about 31 mg / kg, about 32 mg / kg, about 33 mg / kg, about 34 mg / kg, about 35 mg / kg, about 36 mg / kg, about 37 mg / kg, about 38 mg / kg, about 39 mg / kg, about 40 mg / kg, about 41 mg / kg, about 42 mg / kg,about 43 mg / kg, about 44 mg / kg about 45 mg / kg, about 46 mg / kg, about 47 mg / kg, about 48 mg / kg, about 49 mg / kg or about 50 mg / kg, once, twice or three times per week (e.g., 7 days).

[0104] In some embodiments, if pursuant to the above method, it is determined that the subject should not continue to be treated with a glycogen synthase kinase-3 inhibitor, then patient can be switched to treatment with standard of care.

[0105] hr some embodiments, the glycogen synthase kinase-3 inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthase kinase kinase-3alpha inhibitor is administered in combination with one or more chemotherapeutic agents. In some aspects, a glycogen synthase kinase kinase-3beta inhibitor is administered in combination with one or more chemotherapeutic agents. In still other aspects, elraglusib is administered in combination with one or more chemotherapeutic agents.

[0106] In certain embodiments, the chemotherapeutic agents used in the combination is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan / liposomal irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan / liposomal irinotecan leucovorin and fluorouracil. In certain embodiments, the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.EXAMPLESThe following examples are for the purposes of illustration only and are not intended to limit the scope of the claims.EXAMPLE 1Pre-dose plasma samples were obtained from 37 (GnP arm) and 89 (elraglusib / GnP arm) mPDAC patients previously untreated for advanced disease. Mutational analysis of cfDNA extracted from plasma samples was performed by Tempus (Chicago, IL) using the xF-l- platform.EXAMPLE 2In 126 pancreatic cancer patients, frequently mutated genes (>10%) consisted of KRAS (93 / 126, 74%), TP53 (79 / 126, 63%), CDKN2A (26 / 126, 21%), DNMT3A (25 / 126, 20%), NOTCH1 / 2 (25 / 126, 20%), and MLL3 (23 / 126, 18%). Gain-of-function mutations in KRAS, and loss-of- function mutations of TP53, and CDKN2A genes were associated with worse overall survival (OS)in patients treated with elraglusib / GnP (P<0.05) but not in GnP-treated patients. Multivariate analysis of frequently mutated genes showed that patients with double (KRAS+TP53) and triple (KRAS+TP53+CDKN2A) mutations had significantly worse OS only in the elraglusib / GnP arm. The presence of DNMT3A, NOTCH 1 / 2 or MLL3 gene mutations did not correlate with any clinical outcomes.

[0107] It is understood that the foregoing detailed description and accompanying examples are merely illustrative and are not to be taken as limitations upon the scope of the disclosure, which is defined solely by the appended claims and their equivalents.

[0108] Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and may be made without departing from the spirit and scope thereof.

Claims

CLAIMSWe claim:

1. A method of treating a subject suffering from pancreatic cancer, the method comprising the steps of: a. receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, wherein the at least one gene is KRAS, TP53, CDKN2A, or a combination thereof; and b. administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0; or c. not administering the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

2. The method of claim 1, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least two genes.

3. The method of claim 1, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

4. The method of any of claims 1-3, wherein the method comprises: a. administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; b. administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; c. administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; d. administering to the subject a glycogen synthase kinase-3 inhibitor glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; e. administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; f. administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes;g. administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or h. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; i. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; j . not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; k. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; l. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or m. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or n. not administering to the subject a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

5. The method of any of claims 1-4, wherein the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

6. The method of claim 5, wherein the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, or any combinations thereof.

7. The method of claim 5, wherein the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

8. The method of any of claims 1-7, wherein the subject is treated with a combination of a glycogen synthase kinasc-3bcta inhibitor and a chemotherapeutic agent.

9. The method of claim 8, wherein the chemotherapeutic agent is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan, leucovorin and fluorouracil.

10. The method of claim 9, wherein the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

11. The method of any of claims 1-10, wherein the glycogen synthase kinase-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

12. The method according to claim 11, wherein the glycogen synthase kinase-3 inhibitor is elraglusib.

13. The method according to any of claims 1-12, wherein the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

14. The method according to any of claims 1-13, wherein the total number of mutations in the at least one gene is determined using a sequencing assay.

15. A method of selecting a subject suffering from pancreatic cancer for treatment with a glycogen synthase kinase-3 inhibitor, the method comprising the steps of: a. receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer, wherein the at least one gene is KRAS, TP53, CDKN2A or a combination thereof; and b. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0; or c. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

16. The method of claim 15, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least two genes.

17. The method of claim 15, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

18. The method of any of claims 15-17, wherein the method comprises: a. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; b. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; c. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene;d. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; e. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; f. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; g. selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or h. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; i. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; j. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; k. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; l. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or m. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; or n. not selecting a subject for treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

19. The method of any of claims 15-18, wherein the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

20. The method of claim 19, wherein the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, or any combinations thereof.

21. The method of claim 19, wherein the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

22. The method of any of claims 15-21, wherein the subject is treated with a combination of a glycogen synthase kinase-3 inhibitor and a chemotherapeutic agent.

23. The method of claim 22, wherein the chemotherapeutic agent is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan, leucovorin and fluorouracil.

24. The method of claim 23, wherein the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

25. The method of any of claims 15-24, wherein the glycogen synthase kinase-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

26. The method according to claim 25, wherein the glycogen synthase kinase-3 inhibitor is elraglusib.

27. The method according to any of claims 15-26, wherein the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

28. The method according to any of claims 15-27, wherein the total number of mutations in the at least one gene is determined using a sequencing assay.

29. A method of monitoring a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor, the method comprising the steps of: a. receiving information on a total number of mutations in at least one gene in a biological sample obtained from a subject suffering from pancreatic cancer and receiving treatment with a glycogen synthase kinase-3 inhibitor, wherein the at least one gene is KRAS, TP53, CDKN2A or a combination thereof; and b. continuing to treat the subject with the glycogen synthase kinasc-3 inhibitor if the subject has a total number of mutations of 0; or c. discontinuing treatment with the glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 1 or more.

30. The method of claim 29, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least one gene.

31. The method of claim 29, wherein the method comprises receiving information on the total number of mutations in the at least one biological sample in at least three genes.

32. The method of any of claims 29-31, wherein the method comprises: a. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the KRAS gene; b. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the TP53 gene; c. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in the CDKN2A gene; d. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the TP53 genes; e. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS and the CDKN2A genes; f. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the TP53 and the CDKN2A genes; g. continuing to treat the subject with a glycogen synthase kinase-3 inhibitor if the subject has a total number of mutations of 0 in each of the KRAS, TP53 and the CDKN2A genes; or h. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS gene; i. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the in the TP53 gene; j. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the CDKN2A gene; k. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the TP53 genes; l. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS and / or the CDKN2A gene; or m. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or mutations in the TP53 and / or the CDKN2A gene; orn. discontinuing treatment with a glycogen synthase kinase-3 inhibitor if the subject has 1 or more mutations in the KRAS, TP53 and / or CDKN2A genes.

33. The method of any of claims 29-32, wherein the biological sample is a liquid sample, a tumor biopsy sample, or a combination thereof.

34. The method of claim 33, wherein the liquid sample, is a whole blood sample, a plasma sample, a serum sample, a urine sample, or any combinations thereof.

35. The method of claim 33, wherein the tumor biopsy sample is a brush cytology biopsy sample, or a piece of tumor tissue.

36. The method of any of claims 29-35, wherein the subject is treated with a combination of a glycogen synthase kinase-3 inhibitor and a chemotherapeutic agent.

37. The method of claim 36, wherein the chemotherapeutic agent is a taxane, a combination of leucovorin calcium, fluorouracil, irinotecan, and oxaliplatin, gemcitabine, a combination of gemcitabine and taxane, or a combination of irinotecan, leucovorin and fluorouracil.

38. The method of claim 37, wherein the taxane is paclitaxel, nab-paclitaxel, or a combination thereof.

39. The method of any of claims 29-38, wherein the glycogen synthase kinase-3 inhibitor is a glycogen synthase kinase-3alpha inhibitor, a glycogen synthase-3beta inhibitor, or a combination thereof.

40. The method according to claim 39, wherein the glycogen synthase kinase-3 inhibitor is elraglusib.

41. The method according to any of claims 29-40, wherein the method further comprises obtaining a biological sample from a subject suffering from pancreatic cancer and determining the total number of mutations in at least one gene.

42. The method according to any of claims 29-41, wherein the total number of mutations in the at least one gene is determined using a sequencing assay.

Images