Inherited variants in SRD5a genes and response to hormonal therapy in prostate cancer

Abstract

Methods for treating a patient with metastatic castration sensitive prostate cancer that is naive to androgen ablation therapy, comprising administering an effective amount of a non- hormonal therapy alone or in combination with CAB therapy, wherein the patient has polymorphisms in SRD5A genes.

Description

Atty Dkt.: 064189-4890INHERITED VARIANTS IN SRD5A GENES AND RESPONSE TO HORMONAL THERAPY IN PROSTATE CANCERSTATEMENT OF GOVERNMENT SUPPORT

[0001] This invention was made with government support under grant number CA172436, awarded by the National Institutes of Health (NIH) / NCI. The government has certain rights in the invention.BACKGROUND

[0002] Rates of metastatic prostate cancer (mPCa) have been increasing since 2008. It has been projected that by 2025, more than 15,000 new cases of mPCa will be diagnosed each year in the US, an increase of 42% from the year 2015.

[0003] Androgen deprivation therapy (ADT) with luteinizing hormone-releasing hormone (LHRH) agonist or antagonist, along with drugs affecting androgen receptor (AR) signaling (also known as combined androgen blockade (CAB)) is the cornerstone of treatment for metastatic castration-sensitive prostate cancer (mCSPC). Unfortunately, the clinical benefit of this therapy is highly variable, ranging from a few months to many years. Response duration is difficult to predict because little is known about the biological interaction between the hormonal anti-cancer agents, the patient, and the tumor.

[0004] For men with metastatic castration sensitive prostate cancer (mCSPC) androgen deprivation therapy (ADT) in combination with a CYP 17,20 lyse androgen biosynthesis inhibitor or second-generation androgen receptor inhibitors (SGARIs) (combined androgen blockade (CAB)) is the mainstay of treatment1. While initially effective, the duration of benefit from hormonal therapy is highly variable, ranging from a few months to several years, and it is difficult to predict which men will benefit reliably and the duration of response1'3.

[0005] Thus, the current treatment of mCSPC has remained empiric, and the inability to identify distinct patient subgroups early in their clinical course has hindered the development of personalized strategies. As such, a need exists in the art to provide targeted personalized therapies for mCSPC and identify patients that are most effectively treated with current-1-4916-161 1 -5558.1Atty Dkt.: 064189-4890 combination therapies. This disclosure satisfies these needs and provides related advantages as well.SUMMARY OF THE DISCLOSURE

[0006] This disclosure is directed to methods to improve the clinical outcome of patients with mCSPC who may be prescribed hormonal therapies. To that end, this disclosure introduces a novel approach for stratifying and treating patients with metastatic castration-sensitive prostate cancer (mCSPC) based on inherited genetic polymorphisms within the SRD5A gene family (SRD5A1, SRD5A2, SRD5A3), which encode the 5-alpha steroid reductase isoenzymes essential for androgen metabolism.

[0007] The method involves diagnostic and prognostic testing by genotyping patient samples for defined panels of SRD5A variants. Genotyping can be performed using techniques such as PCR, nucleic acid hybridization, Illumina arrays, whole genome sequencing, or streamlined single SNP assays, including detection of the SRD5A2 rs632148 variant.

[0008] Therapeutic decisions are guided by the patient’s SRD5A genotype. Patients who carry certain SRD5 A variants are identified as poor responders to conventional androgen deprivation therapy (ADT) and androgen receptor pathway inhibitors, particularly CYP 17,20 lyase inhibitors (e.g., abiraterone acetate). For these individuals, the method recommends upfront non-hormonal treatments, such as chemotherapy, immunotherapy, or PARP inhibitors.

[0009] Clinical validation of this approach is provided by large-scale data, including results from the SWOG S1216 trial, which demonstrate that carriers of SRD5A variants exhibit significantly reduced progression-free survival, overall survival, and PSA responses to hormonal therapies. These findings show a clear dose-response relationship and biological plausibility, distinguishing this method from previous studies.

[0010] The application details the identification and validation of a prognostic genetic panel, incorporating multiple genes and SNPs within the SRD5 A family, or a representative proxy SNP like rs632148. This panel captures functionally significant variations, enabling robust prediction of therapy response.-2-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0011] Importantly, the application provides a personalized therapeutic algorithm that uses SRD5 A genotype data to prospectively select the most appropriate first-line therapy — hormonal or non-hormonal — for individual patients. To the best of the Applicant’s knowledge, this actionable decision-making process, guided by genetic biomarkers, is not taught or suggested by existing approaches, marking a shift from empirical to biomarker- driven prostate cancer management.

[0012] The clinical utility of this method is supported by rigorous, multicenter, prospective clinical trial data that confirm the mechanism of action and outcome associations, addressing the limitations and lack of plausibility in previous single-SNP or population-limited studies.

[0013] To facilitate implementation, the method simplifies complex genomic data into actionable diagnostics, such as single-SNP assays using TaqMan® technology or whole genome sequencing, making it highly practical for clinical use.

[0014] Ultimately, this disclosure provides the explicit integration of SRD5A genotype- guided therapy by linking genetic results to the initial selection of non-hormonal treatments for poor responders. This use of genetic data to directly inform therapeutic decisions constitutes an inventive and clinically meaningful workflow.

[0015] As explained in more detail herein, steroid 5a-reductase isoenzymes, encoded by the SRD5A1, SRD5A2, and SRD5A3 genes, catalyze the conversion of testosterone to the more potent androgen, 5a-dihydrotestosterone (DHT). Without being bound by theory, Applicant hypothesized that inherited variants in SRD5 A genes may influence DHT availability, especially under castrate testosterone levels produced by androgen deprivation therapy (ADT) and thus may potentially impact disease progression.

[0016] In one embodiment, Applicant studied men with metastatic castration sensitive prostate cancer enrolled in SWOG S1216, a phase III randomized multicenter clinical trial of Androgen deprivation therapy (ADT) with a CYP 17,20 lyse inhibitor (TAK-700) or an androgen receptor inhibitor (bicalutamide). Inherited variants (17 variants in SRD5A1, 19 in SRD5A2, and 5 in SRD5A3) were assayed using the Illumina Global Diversity Array. Associations with progression free survival (PFS) and overall survival (OS) were analyzed using Kaplan-Meier curves and Cox proportional hazards models. Associations with 7- month PSA response (< 0.2, 0.2-4.0, >4.0 ng / ml) were analyzed using ordinal Chi-Square-3-4916-161 1 -5558.1Atty Dkt.: 064189-4890 tests and multinomial logistic regression models. All models were adjusted for treatment arm, extent of disease at study entry (extensive v. minimal), Zubrod performance status, and ADT status at baseline (already initiated or not).

[0017] Of 364 men with genotype data, 38 (10.4%) carried variant alleles on one or more of the SRD5A genes and 25 (66%) of these men carried variant alleles on all three genes.Variant allele carriers had shorter PFS (HR=3.6; 95% CI 2.5-5.3; p<0.001) and were less likely to experience complete PSA response to treatment (p=0.006). For OS there was a statistically significant interaction between treatment arm and variant allele status (p=0.02). Among those in the TAK-700 arm, median OS was 25.5 months for variant carriers vs. 94.7 months for non-carriers (HR=5.4; 95% CI 2.9-10.1; p<0.001). The association was weaker in the bicalutamide arm: median OS was 39.7 months for variant carriers vs. 86.1 months for non-carriers (HR=1.9; 95% CI 1.1-3.1; p=0.02).

[0018] Men carrying SRD5A gene variants are more likely to experience worse outcomes in response to combined androgen blockade (CAB) therapy, especially when ADT is combined with a selective CYP 17,20 lyase inhibitor.

[0019] Thus, this disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising, or consisting essentially of, or yet further consisting of, administering an effective amount of a non-hormonal therapy, wherein the patient has an inherent variant genotype for a SRD5A gene.

[0020] In some aspects, the disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising, or consisting essentially of, or yet further consisting of, administering an effective amount of a hormonal therapy, wherein the patient lacks an inherent variant genotype for a SRD5A gene.

[0021] In some aspects, the disclosure provides a method for determine whether a prostate cancer patient who is naive to androgen ablation therapy is likely to be responsive to non- hormonal therapy, the method comprising, or consisting essentially of, or yet further consisting of, determining whether a prostate tumor sample isolated from the patient contains a an inherent variant polymorphism in an SRD5A gene, wherein the patient is more likely to-4-4916-161 1 -5558.1Atty Dkt.: 064189-4890 be responsive to non-hormonal therapy when the patient has the inherent variant polymorphism in the SRD5A gene and is less likely to be responsive to non-hormonal therapy when the patient lacks the inherent variant polymorphism or has wildtype genotype the SRD5A gene.BRIEF DESCRIPTION OF THE FIGURES

[0022] FIGS. 1A - IB: Association of SRD5A variant status with overall survival, stratified by treatment arm. FIG. 1A shows overall survival / TAK-700. FIG. IB shows overall survival / Bicalutamide.

[0023] FIG. 2: SRD5A variant allele burden among variant allele carriers (N=38). A total of 31 SRD5A variants were assayed (17 in SRD5A1, 19 in SRD5A2, and 5 in SRD5A3) on the Illumina Global Diversity Array. All variant allele carriers tested positive for 4 or more variants. The median number of variants carried was 15, and two of the men carried 22 of the 31 variant alleles.

[0024] FIGS. 3A - 3C: Association of SRD5A variants with progression free survival (Log-rank test). For SRD5A1 (FIG. 3A), 15 of the 17 SNPs were associated with progression free survival with p<0.01, and 9 of the variants had p values less than 10'5(i.e. negative logio(p value) greater than 5. Results were similar for SRD5A2 (FIG. 3B), with 12 of 19 SNPs significant at the 0.01 level, and 10 of those significant with p<10'5. For SRD5A3 (FIG. 3C), 4 of the 5 SNPs were significant at the 0.01 level, with 2 of those significant with p<10'5.

[0025] FIG. 4: Association of SRD5A variant status with progression free survival.Variant allele carriers had shorter progression free survival than non-carriers (HR=3.6; 95% CI 2.5-5.3; p<0.001). Median progression free survival was 9.9 months for variant carriers vs. 41.0 months for non-carriers.

[0026] FIGS. 5A - 5B: Association of SRD5A variant status with overall survival, stratified by treatment arm. Among those in the TAK-700 arm (ADT plus a CYP17,20 lyase inhibitor), median overall survival was 25.5 months for variant carriers vs. 94.7 months for non-carriers (HR=5.4; 95% CI 2.9-10.1; p<0.001). The association was weaker in the Bicalutamide arm (ADT plus an androgen receptor inhibitor). Median overall survival was-5-4916-161 1 -5558.1Atty Dkt.: 064189-489039.7 months for variant carriers vs. 86.1 months for non-carriers (HR=1.9; 95% CI 1.1 -3.1; p=0.02).

[0027] FIGS. 6A and 6B: Progression free survival and overall survival by SRD5A variant carrier status. FIG. 6A, dashed lines represent SRD5 A variant carriers; Solid lines represent non-carriers. In FIG. 6B, black lines represent the Taq700 arm (ADT plus a CYP 17,20 lyse inhibitor); gray lines represent the Bicalutamide arm (ADT plus a first- generation androgen receptor inhibitor).DETAILED DESCRIPTION

[0028] Definitions

[0029] Throughout this disclosure, various publications, patents and published patent specifications are referenced by an identifying citation or by an Arabic number referencing a citation found immediately preceding the claims. The disclosures of these publications, patents and published patent specifications are hereby incorporated by reference into the present disclosure to more fully describe the state of the art to which this disclosure pertains.

[0030] As used herein, certain terms may have the following defined meanings. As used in the specification and claims, the singular form “a,” “an” and “the” include singular and plural references unless the context clearly dictates otherwise. For example, the term “a cell” includes a single cell as well as a plurality of cells, including mixtures thereof.

[0031] As used herein, the term “comprising” is intended to mean that the methods include the recited elements, but not excluding others. “Consisting essentially of’ when used to define methods, shall mean excluding other elements of any essential significance to the method. “Consisting of’ shall mean excluding more than trace elements of other ingredients for claimed compositions and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this disclosure. Accordingly, it is intended that the methods can include additional steps and components (comprising) or alternatively including steps of no significance (consisting essentially of) or alternatively, intending only the stated method steps (consisting of).

[0032] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which are varied ( + ) or ( - ) by increments of-6-4916-161 1 -5558.1Atty Dkt.: 064189-48900.1. It is to be understood, although not always explicitly stated that all numerical designations are preceded by the term “about”. The term “about” also includes the exact value “X” in addition to minor increments of “X” such as “X + 0.1” or “X - 0.1.” It also is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0033] The term “therapy” encompasses cancer therapies that employ chemical or biological agents or other therapies, such as radiation therapies, radionucleotide therapy, a small molecule drug or a large molecule, such as antibodies, CAR therapies, RNAi and gene therapies. Non-limiting examples of chemotherapies are provided below. Unless specifically excluded, when a specific therapy is recited, equivalents of the therapy are within the scope of this disclosure.

[0034] An “effective amount” is an amount sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages. Such delivery is dependent on a number of variables including the time period for which the individual dosage unit is to be used, the bioavailability of the therapeutic agent, the route of administration, etc. It is understood, however, that specific dose levels of the therapeutic agents disclosed herein for any particular subject depends upon a variety of factors including the activity of the specific compound employed, bioavailability of the compound, the route of administration, the age of the animal and its body weight, general health, sex, the diet of the animal, the time of administration, the rate of excretion, the drug combination, and the severity of the particular disorder being treated and form of administration. In general, one will desire to administer an amount of the compound that is effective to achieve a serum level commensurate with the concentrations found to be effective in vivo. These considerations, as well as effective formulations and administration procedures are well known in the art and are described in standard textbooks.

[0035] As used herein, “treating” or “treatment” of a disease in a subject refers to (1) preventing the symptoms or disease from occurring in a subject that is predisposed or does not yet display symptoms of the disease; (2) inhibiting the disease or arresting its development; or (3) ameliorating or causing regression of the disease or the symptoms of the disease. As understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of this technology, beneficial or -7-4916-161 1 -5558.1Atty Dkt.: 064189-4890 desired results can include one or more, but are not limited to, alleviation or amelioration of one or more symptoms, diminishment of extent of a condition (including a disease), stabilized (i.e., not worsening) state of a condition (including disease), delay or slowing of condition (including disease), progression, amelioration or palliation of the condition (including disease), states and remission (whether partial or total), whether detectable or undetectable. In one aspect, treatment excludes prophylaxis.

[0036] When the disease is cancer, the following clinical endpoints are non-limiting examples of treatment: (1) elimination of a cancer in a subject or in a tissue / organ of the subject or in a cancer loci; (2) reduction in tumor burden (such as number of cancer cells, number of cancer foci, number of cancer cells in a foci, size of a solid cancer, concentrate of a liquid cancer in the body fluid, and / or amount of cancer in the body); (3) stabilizing or delay or slowing or inhibition of cancer growth and / or development, including but not limited to, cancer cell growth and / or division, size growth of a solid tumor or a cancer loci, cancer progression, and / or metastasis (such as time to form a new metastasis, number of total metastases, size of a metastasis, as well as variety of the tissues / organs to house metastatic cells); (4) less risk of having a cancer growth and / or development; (5) inducing an immune response of the patient to the cancer, such as higher number of tumor-infiltrating immune cell, higher number of activated immune cells, or higher number cancer cell expressing an immunotherapy target, or higher level of expression of an immunotherapy target in a cancer cell; (6) higher probability of survival and / or increased duration of survival, such as increased overall survival (OS, which may be shown as 1-year, 2-year, 5-year, 10-year, or 20-year survival rate), increased progression free survival (PFS), increased disease free survival (DFS), increased time to tumor recurrence (TTR) and increased time to tumor progression (TTP). In some embodiments, the subject after treatment experiences one or more endpoints selected from tumor response, reduction in tumor size, reduction in tumor burden, increase in overall survival, increase in progression free survival, inhibiting metastasis, improvement of quality of life, minimization of drug-related toxicity, and avoidance of side-effects (e.g., decreased treatment emergent adverse events). In some embodiments, improvement of quality of life includes resolution or improvement of cancer-specific symptoms, such as but not limited to fatigue, pain, nausea / vomiting, lack of appetite, and constipation; improvement or maintenance of psychological well-being (e.g., degree of irritability, depression, memory-8-4916-161 1 -5558.1Atty Dkt.: 064189-4890 loss, tension, and anxiety); improvement or maintenance of social well-being (e.g., decreased requirement for assistance with eating, dressing, or using the restroom; improvement or maintenance of ability to perform normal leisure activities, hobbies, or social activities; improvement or maintenance of relationships with family). In some embodiments, improved patient quality of life that is measured qualitatively through patient narratives or quantitatively using validated quality of life tools known to those skilled in the art, or a combination thereof. Additional non-limiting examples of endpoints include reduced hospital admissions, reduced drug use to treat side effects, longer periods off-treatment, and earlier return to work or caring responsibilities. In one aspect, prevention or prophylaxis is excluded from treatment.

[0037] The term “subject,” “host,” “individual,” and “patient” are as used interchangeably herein to refer to animals, typically mammalian animals. Any suitable mammal can be treated by a method described herein. Non-limiting examples of mammals include humans, non-human primates (e.g., apes, gibbons, chimpanzees, orangutans, monkeys, macaques, and the like), domestic animals (e.g., dogs and cats), farm animals (e.g., horses, cows, goats, sheep, pigs) and experimental animals (e.g., mouse, rat, rabbit, guinea pig). In some embodiments, a mammal is a human. A mammal can be any age or at any stage of development (e.g., an adult, teen, child, infant, or a mammal in utero). A mammal can be male or female. In some embodiments, a subject is a human. In some embodiments, a subject has or is diagnosed of having or is suspected of having a cancer.

[0038] A cancer may refer to a local cancer (which is an invasive malignant cancer confined entirely to the organ or tissue where the cancer began), a metastatic cancer (referring to a cancer that spreads from its site of origin to another part of the body), a non-metastatic cancer, a primary cancer (a term used describing an initial cancer a subject experiences), a secondary cancer (referring to a metastasis from primary cancer or second cancer unrelated to the original cancer), an advanced cancer, an unresectable cancer, or a recurrent cancer. As used herein, an advanced cancer refers to a cancer that had progressed after receiving one or more of: the first line therapy, the second line therapy, or the third line therapy.

[0039] “ Castrate-sensitive prostate cancer” is prostate cancer that needs androgens (male hormones) to grow and therefore stops growing when androgens are not present. Many early-stage prostate cancers are androgen-dependent, so reducing the amount of androgens in-9-4916-161 1 -5558.1Atty Dkt.: 064189-4890 the body or blocking their action may be an effective type of therapy. Also called androgendependent prostate cancer, androgen-sensitive prostate cancer, CSPC, hormone-sensitive prostate cancer (HSPC). See https: / / www.cancer.gov / publications / dictionaries / cancer- terms / def / castrate-sensitive-prostate-cancer, last accessed on August 24, 2025.

[0040] “ Testosterone suppressive therapy” or “androgen suppression” is a medical treatment to suppress or block the production or action of male sex hormones, typically used against certain cancers that rely on male growth hormones. This a done by castration (sometimes chemical castration or by prescribing female sex hormones and antiandrogen drugs.

[0041] As used herein, “androgen deprivation therapy” is (ADT) refers to any medical, chemical, or surgical intervention that reduces or suppresses the production, activity, or effects of androgens (male hormones such as testosterone and dihydrotestosterone) in the body. ADT commonly includes the administration of luteinizing hormone-releasing hormone (LHRH) agonists or antagonists, gonadotropin-releasing hormone (GnRH) agonists or antagonists, surgical orchiectomy, or other agents that directly or indirectly lower androgen levels or inhibit their action. ADT may be administered alone or in combination with other agents but, in the context of this application, specifically excludes treatments whose principal mechanism is not reduction of androgen signaling.

[0042] As used herein, “non-hormonal therapy” refers to any therapeutic treatment or modality for prostate cancer that does not primarily function by directly suppressing, blocking, or modulating testosterone or androgen receptor signaling. In the context of this application, non-hormonal therapy includes, but is not limited to, chemotherapy (such as docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or pemetrexed / alimta), immunotherapy (such as immune checkpoint inhibitors, CAR-T cell therapy, cancer vaccines, or cytokine therapy), radionucleotide therapy (such as administration of radioisotopes including radium-223 or lutetium-177-labeled ligands), antibody-drug conjugates (monoclonal antibodies conjugated with cytotoxic agents), and poly(ADP -ribose) polymerase (PARP) inhibitors. Non-hormonal therapy explicitly excludes androgen deprivation therapy (ADT), CYP 17,20 lyase inhibitors, androgen receptor inhibitors, and any treatment whose primary mode of action is hormonal manipulation or disruption of androgen signaling.-10-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0043] A “CYP 17,20 lysase inhibitor” is an orally available non-steroidal, lyase-selective inhibitor of the steroid 17-alpha-hydroxylase / C 17,20 lyase (CYP17A1 or CYP17), with potential anti -androgenic and antineoplastic activities. Upon oral administration, CYP 17 lyase inhibitor ASN001 selectively binds to and inhibits the lyase activity of CYP17A1 in both the testes and adrenal glands, resulting in a significant reduction in androgen production to castrate-range levels. This may both decrease androgen-dependent growth signaling and inhibit the proliferation of androgen-dependent tumor cells. Non-limiting examples of such are described below.

[0044] Table 1

[0045] As used herein, “combined androgen blockade” (CAB) therapy refers to a therapeutic regimen for prostate cancer comprising the combination of androgen deprivation therapy (ADT) and an agent that inhibits androgen receptor signaling or activity. For the purposes of this application, CAB therapy specifically excludes any treatment regimen or agent that does not include both an androgen deprivation component and a direct androgen receptor pathway inhibitor or androgen biosynthesis inhibitor as provided above.

[0046] CAB therapy may include, but is not limited to, the concurrent or sequential use of one or more of the following commercially available agents:

[0047] 1. Androgen Deprivation Therapy (ADT) Agents

[0048] LHRH / GnRH agonists: leuprolide acetate (Lupron®, Eligard®), goserelin acetate (Zoladex®), triptorelin pamoate (Trelstar®), histrelin acetate (Vantas®)

[0049] LHRH / GnRH antagonists: degarelix (Firmagon®), relugolix (Orgovyx®)-11-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0050] Surgical castration: orchiectomy

[0051] 2. Androgen Receptor Pathway Inhibitors and Androgen Biosynthesis Inhibitors

[0052] Nonsteroidal antiandrogens: bicalutamide (Casodex®), flutamide (Eulexin®), nilutamide (Nilandron®), enzalutamide (Xtandi®), apalutamide (Erleada®), darolutamide (Nubeqa®)

[0053] Steroidal antiandrogens: cyproterone acetate (Androcur® and generics; primarily in Europe, Canada, and Asia)

[0054] Androgen biosynthesis inhibitors: abiraterone acetate (Zytiga®), administered in combination with prednisone

[0055] 3. Representative CAB Regimens

[0056] Typical commercial CAB therapy includes, for example:

[0057] Leuprolide or goserelin administered with bicalutamide, flutamide, or nilutamide

[0058] Leuprolide, goserelin, triptorelin, or degarelix in combination with enzalutamide, apalutamide, darolutamide, or abiraterone acetate plus prednisone

[0059] Where available, surgical orchiectomy may be substituted for LHRH / GnRH analogue or antagonist therapy.

[0060] Biraterone, enzalutamide, apalutamide, and darolutamide are considered “nextgeneration” agents and are widely used as part of CAB in first- and second-line metastatic prostate cancer therapy.

[0061] Table 2-12-4916-161 1 -5558.1Atty Dkt.: 064189-4890Androgen Biosynthesis InhibitorAbiraterone acetate

[0062] Combined androgen blockade (CAB) is defined as androgen deprivation therapy (ADT) with luteinizing hormone-releasing hormone (LHRH) agonist or antagonist, along with drugs affecting androgen receptor (AR) signaling in combination with CYP17 or AR inhibitors (ARIs). Non-limiting examples of androgen receptor signaling inhibitors (ARSIs), include for example, enzalutamide, apalutamide, darolutamide, and abiraterone.

[0063] Androgen receptor signaling inhibitor” means any agent that directly impedes androgen receptor activity or prevents androgen-mediated signaling in prostate cancer cells, including but not limited to nonsteroidal and steroidal antiandrogens such as bicalutamide, enzalutamide, apalutamide, darolutamide, and cyproterone acetate.

[0064] Enzalutamide is a nonsteroidal androgen receptor inhibitor used as an oral antiandrogen drug for the treatment of prostate cancer. It binds to the androgen receptor and prevents androgen-induced signaling, thereby inhibiting the growth of androgen-dependent prostate cancer cells. Enzalutamide is marketed under the trade name Xtandi®.

[0065] Apalutamide is a nonsteroidal androgen receptor antagonist administered orally for the treatment of prostate cancer. It blocks binding of androgens to the androgen receptor, impeding androgen receptor signaling and suppressing the proliferation of prostate cancer cells. Apalutamide is marketed under the trade name Erleada®.

[0066] Darolutamide is a nonsteroidal anti-androgen drug used in the treatment of prostate cancer. It inhibits androgen receptor signaling by directly binding to the androgen receptor, thereby blocking the growth-promoting effects of androgens. Darolutamide is administered orally and is sold under the trade name Nubeqa®.

[0067] Abiraterone is a selective androgen biosynthesis inhibitor indicated for the treatment of prostate cancer. It irreversibly inhibits the enzyme CYP17A1 (17a-hydroxylase / C17,20- lyase), which is required for androgen production in the testes, adrenal glands, and prostate tumor tissue. Abiraterone is usually administered in combination with prednisone and is marketed under the trade name Zytiga®.

[0068] As used herein, “an inherent variant genotype for a SRD5 A gene” refers to a germline (inherited) genetic variant in at least one member of the steroid 5 alpha-reductase gene-13-4916-161 1 -5558.1Atty Dkt.: 064189-4890 family, which includes SRD5A1, SRD5A2, and SRD5A3. An inherent variant genotype is distinct from the wild-type (reference) genotype and may comprise one or more of the following types of genetic alterations: single nucleotide polymorphism (SNP), small insertion, small deletion, or other inherited sequence variation located at a defined genomic position (such as an rs number). These variants are present in all nucleated cells of an individual from birth and can be detected in genomic DNA isolated from blood, tissue, or other biological samples. In one aspect, an “inherent variant genotype for a SRD5A gene” encompasses, but is not limited to, any of the following SNPs:

[0069] SRD5A1 : rs477930, rs8192139, rs535981, rs562461, rs248805, rs3822430, rs8192206, rs4702381, rs8192249, rs3297, rsl042150, rsl6877779, rsl2716180, rs79421055, rs28594474, rsl651071, rs2434525;

[0070] SRD5A2: rs28383087, rs28383086, rs28383085, rs9332975, rs28383083, rsl92604242, rs28383082, rsl2470143, rs2366063, rs6727380, rs551741548, rs72794634, rs781063043, rs2268796, rs2300697, rsl988909, rs74403699, rs632148, rs559555;

[0071] SRD5A3: rs73236134, rs73236133, rs7674388, rs7664100, rs7663650.

[0072] The term also encompasses functionally equivalent or clinically relevant germline variants of SRD5A1, SRD5A2, or SRD5A3 not explicitly listed above, provided such variants are inherited and not acquired somatically within tumor tissue.Detection of an inherent variant genotype can be accomplished by PCR, sequencing, microarray analysis, TaqMan® SNP assay, or other nucleic acid-based genotyping methods.

[0073] The term “contacting” means direct or indirect binding or interaction between two or more. A particular example of direct interaction is binding. A particular example of an indirect interaction is where one entity acts upon an intermediary molecule, which in turn acts upon the second referenced entity. Contacting as used herein includes in solution, in solid phase, in vitro, ex vivo, in a cell and in vivo. Contacting in vivo can be referred to as administering, or administration.

[0074] As used herein, the term “administration” and “administering” are used to mean introducing an agent into a subject. Routes of administration include, but are not limited to, oral (such as a tablet, capsule or suspension), topical, transdermal, intranasal, vaginal, rectal, subcutaneous intravenous, intravenous, intraarterial, intramuscular, intraosseous,-14-4916-161 1 -5558.1Atty Dkt.: 064189-4890 intraperitoneal, intraocular, subconjunctival, sub-Tenon’s, intravitreal, retrobulbar, intracameral, intratumoral, epidural and intrathecal. The mode of administration will differ with the patient being treated, as determined by the treating physician.

[0075] An “immunotherapy agent” means a type of cancer treatment which uses a patient’s own immune system to fight cancer, including but not limited to a physical intervene, a chemical substance, a biological molecule or particle, a cell, a tissue or organ, or any combinations thereof, enhancing or activating or initiating a patient’s immune response against cancer. Non-limiting examples of immunotherapy agents include antibodies, immune regulators, checkpoint inhibitors, an antisense oligonucleotide (ASO), a RNA interference (RNAi), a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) system, a viral vector, an anti-cancer cell therapy (e.g., transplanting an anti-cancer immune cell optionally amplified and / or activated in vivo, or administering an immune cell expressing a chimeric antigen receptor (CAR)), a CAR therapy, and cancer vaccines.

[0076] As used herein, immune checkpoint refers to a regulator and / or modulator of the immune system (such as an immune response, an anti-tumor immune response, a nascent anti-tumor immune response, an anti-tumor immune cell response, an anti-tumor T cell response, and / or an antigen recognition of T cell receptor in the process of immune response). Their interaction activates either inhibitory or activating immune signaling pathways. Thus a checkpoint may contain one of the two signals: an stimulatory immune checkpoint that stimulates an immune response, and an inhibitory immune checkpoint inhibiting an immune response. In some embodiments, the immune checkpoint is crucial for self-tolerance, which prevents the immune system from attacking cells indiscriminately. However, some cancers can protect themselves from attack by stimulating immune checkpoint targets. In some embodiments, the immune checkpoints are present on T cells, antigen-presenting cells (APCs) and / or tumor cells.

[0077] One target of an immunotherapy agent is a tumor-specific antigen while the immunotherapy directs or enhances the immune system to recognize and attack tumor cells. Non-limiting examples of such agent includes a cancer vaccine presenting a tumor-specific antigen to the patient’s immune system, a monoclonal antibody or an antibody-drug conjugate specifically binding to a tumor-specific antigen, a bispecific antibody specifically binding to a tumor-specific antigen and an immune cell (such as a T-cell engager or a NK- -15-4916-161 1 -5558.1Atty Dkt.: 064189-4890 cell engager), an immune cell (such as a killer cell) specifically binding to a tumor-specific antigen (such as a CAR-T cell, a CAR-NK cell, and a CAR-NKT cell), a polynucleotide (or a vector comprising the same) transfecting / transducing an immune cell to express an tumorspecific antibody of an antigen binding fragment thereof (such as a CAR), or a polynucleotide (or a vector comprising the same) transfecting / transducing a cancer cell to express an antigen or a marker which can be recognized by an immune cell.

[0078] Another exemplified target is an inhibitory immune checkpoint which suppresses the nascent anti-tumor immune response, such as A2AR, B7-H3, B7-H4, BTLA, CTLA-4, CTLA-4 / B7-1 / B7-2, IDO, KIR, LAG3, N0X2, PD-1, PD-L1 and TIM-3, VISTA, SIGLEC7 (Sialic acid-binding immunoglobulin-type lectin 7, also designated as CD328) and SIGLEC9 (Sialic acid-binding immunoglobulin-type lectin 9, also designated as CD329). Non-limiting examples of such agent includes an antagonist or inhibitor of an inhibitory immune checkpoint, an agent reducing the expression and / or activity of an inhibitory immune checkpoint (such as via an antisense oligonucleotide (ASO), a RNA interference (RNAi), or a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) system), an antibody or an antibody-drug conjugate or a ligand specifically binding to and reducing (or inhibiting) the activity of an inhibitory immune checkpoint, an immune cell with reduced (or inhibited) an inhibitory immune checkpoint (and optionally specifically binding to a tumor-specific antigen, such as a CAR-T cell, a CAR-NK cell, and a CAR-NKT cell), and a polynucleotide (or a vector comprising the same) transfecting / transducing an immune cell or a cancer cell to reduce or inhibit an inhibitory immune checkpoint thereof. Reducing expression or activity of such inhibitory immune checkpoint enhances immune response of a patient to a cancer.

[0079] A further possible immunotherapy target is a stimulatory checkpoint molecule (including but not limited to 4-1BB, CD27, CD28, CD40, CD122, CD137, 0X40, GITR and ICOS), wherein the immunotherapy agent actives or enhances the anti-tumor immune response. Non-limiting examples of such agent includes an agonist of a stimulatory checkpoint, an agent increasing the expression and / or activity of a stimulating immune checkpoint, an antibody or an antibody-drug conjugate or a ligand specifically binding to and activating or enhancing the activity of a stimulating immune checkpoint, an immune cell with increased expression and / or activity of a stimulating immune checkpoint (and optionally specifically binding to a tumor-specific antigen, such as a CAR-T cell, a CAR-NK cell, and a-16-4916-161 1 -5558.1Atty Dkt.: 064189-4890CAR-NKT cell), and a polynucleotide (or a vector comprising the same) transfecting / transducing an immune cell or a cancer cell to express a stimulating immune checkpoint thereof.

[0080] In some embodiments, an immunotherapy agent utilizes one or more targets, such as a bispecific T cell engager, a bispecific NK cell engager, or a CAR cell therapy. In some embodiments, the immunotherapy agent targets one or more immune regulatory or effector cells.

[0081] The term “chimeric antigen receptor” (CAR), as used herein, refers to a fused protein comprising an extracellular domain capable of binding to an antigen, a transmembrane domain derived from a polypeptide different from a polypeptide from which the extracellular domain is derived, and at least one intracellular domain. The “chimeric antigen receptor (CAR)” is sometimes called a “chimeric receptor”, a “T-body”, or a “chimeric immune receptor (CIR).” The “extracellular domain capable of binding to an antigen” means any oligopeptide or polypeptide that can bind to a certain antigen. The “intracellular domain” or “intracellular signaling domain” means any oligopeptide or polypeptide known to function as a domain that transmits a signal to cause activation or inhibition of a biological process in a cell. In certain embodiments, the intracellular domain may comprise, alternatively consist essentially of, or yet further comprise one or more costimulatory signaling domains in addition to the primary signaling domain. The “transmembrane domain” means any oligopeptide or polypeptide known to span the cell membrane and that can function to link the extracellular and signaling domains. A chimeric antigen receptor may optionally comprise a “hinge domain” which serves as a linker between the extracellular and transmembrane domains.

[0082] As used herein, a CAR therapy may refer to administrating an immune cell expressing a CAR into a subject as well as contacting a vector expressing a CAR in an immune cell (such as in vivo).

[0083] As used herein, the term “NK cell,” also known as natural killer cell, refers to a type of lymphocyte that originates in the bone marrow and play a critical role in the innate immune system. NK cells provide rapid immune responses against viral-infected cells, tumor cells or other stressed cell, even in the absence of antibodies and major histocompatibility-17-4916-161 1 -5558.1Atty Dkt.: 064189-4890 complex on the cell surfaces. NK cells for using in a cell therapy and / or a CAR therapy may either be isolated or obtained from a commercially available source.

[0084] The phrase “first line” or “second line” or “third line” refers to the order of treatment received by a patient. First line therapy regimens are treatments given first, whereas second or third line therapy are given after the first line therapy or after the second line therapy, respectively. The National Cancer Institute defines first line therapy as “the first treatment for a disease or condition. In patients with cancer, primary treatment can be surgery, chemotherapy, radiation therapy, or a combination of these therapies. First line therapy is also referred to those skilled in the art as “primary therapy and primary treatment.” See National Cancer Institute website at cancer.gov. Typically, a patient is given a subsequent chemotherapy regimen because the patient did not show a positive clinical or sub-clinical response to the first line therapy or the first line therapy has stopped.

[0085] In one aspect, the term “equivalent” or “biological equivalent” of an antibody means the ability of the antibody to selectively bind its epitope protein or fragment thereof as measured by ELISA or other suitable methods. Biologically equivalent antibodies include, but are not limited to, those antibodies, peptides, antibody fragments, antibody variant, antibody derivative and antibody mimetics that bind to the same epitope as the reference antibody.

[0086] In one aspect, the term “equivalent” of “chemical equivalent” of a chemical means the ability of the chemical to selectively interact with its target protein, DNA, RNA or fragment thereof as measured by the inactivation of the target protein, incorporation of the chemical into the DNA or RNA or other suitable methods. Chemical equivalents include, but are not limited to, those agents with the same or similar biological activity and include, without limitation a pharmaceutically acceptable salt or mixtures thereof that interact with and / or inactivate the same target protein, DNA, or RNA as the reference chemical.

[0087] The term “allele,” which is used interchangeably herein with “allelic variant” refers to alternative forms of a gene or portions thereof. Alleles occupy the same locus or position on homologous chromosomes. When a subject has two identical alleles of a gene, the subject is said to be homozygous for the gene or allele. When a subject has two different alleles of a gene, the subject is said to be heterozygous for the gene. Alleles of a specific gene can differ-18-4916-161 1 -5558.1Atty Dkt.: 064189-4890 from each other in a single nucleotide, or several nucleotides, and can include substitutions, deletions and insertions of nucleotides. An allele of a gene can also be a form of a gene containing a mutation.

[0088] As used herein, the term “determining the genotype of a cell or tissue sample” intends to identify the genotypes of polymorphic loci of interest in the cell or tissue sample. In one aspect, a polymorphic locus is a single nucleotide polymorphic (SNP) locus. If the allelic composition of a SNP locus is heterozygous, the genotype of the SNP locus will be identified as “X / Y” wherein X and Y are two different nucleotides. If the allelic composition of a SNP locus is heterozygous, the genotype of the SNP locus will be identified as “X / X” wherein X identifies the nucleotide that is present at both alleles.

[0089] The term “genetic marker” refers to an allelic variant of a polymorphic region of a gene of interest and / or the expression level of a gene of interest.

[0090] The term “polymorphism” refers to the coexistence of more than one form of a gene or portion thereof. A portion of a gene of which there are at least two different forms, i.e., two different nucleotide sequences, is referred to as a “polymorphic region of a gene.” A polymorphic region can be a single nucleotide, the identity of which differs in different alleles.

[0091] The term “genotype” refers to the specific allelic composition of an entire cell or a certain gene and in some aspects a specific polymorphism associated with that gene, whereas the term “phenotype” refers to the detectable outward manifestations of a specific genotype.

[0092] The term “isolated” as used herein refers to molecules or biological or cellular materials being substantially free from other materials. In one aspect, the term “isolated” refers to nucleic acid, such as DNA or RNA, or protein or polypeptide, or cell or cellular organelle, or tissue or organ, separated from other DNAs or RNAs, or proteins or polypeptides, or cells or cellular organelles, or tissues or organs, respectively, that are present in the natural source. The term “isolated” also refers to a nucleic acid or peptide that is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. Moreover, an “isolated nucleic acid” is meant to include nucleic acid fragments which are not naturally occurring as fragments and would not be found in the natural state.-19-4916-161 1 -5558.1Atty Dkt.: 064189-4890The term “isolated” is also used herein to refer to polypeptides which are isolated from other cellular proteins and is meant to encompass both purified and recombinant polypeptides. The term “isolated” is also used herein to refer to cells or tissues that are isolated from other cells or tissues and is meant to encompass both cultured and engineered cells or tissues.

[0093] The term “treating” as used herein is intended to encompass curing as well as ameliorating at least one symptom of the condition or disease. For example, in the case of cancer, a response to treatment includes a reduction in cachexia, increase in survival time, elongation in time to tumor progression, reduction in tumor mass, reduction in tumor burden and / or a prolongation in time to tumor metastasis, time to tumor recurrence, tumor response, complete response, partial response, stable disease, progressive disease, progression free survival, overall survival, each as measured by standards set by the National Cancer Institute and the U.S. Food and Drug Administration for the approval of new drugs. In one aspect, treatment excludes prevention or prophylaxis.

[0094] The term “clinical outcome”, “clinical parameter”, “clinical response”, or “clinical endpoint” refers to any clinical observation or measurement relating to a patient’s reaction to a therapy. Non-limiting examples of clinical outcomes include tumor response (TR), overall survival (OS), progression free survival (PFS), disease free survival, time to tumor recurrence (TTR), time to tumor progression (TTP), relative risk (RR), toxicity or side effect.

[0095] The term “suitable for a therapy” or “suitably treated with a therapy” shall mean that the patient is likely to exhibit one or more desirable clinical outcomes as compared to patients having the same disease and receiving the same therapy but possessing a different characteristic that is under consideration for the purpose of the comparison. In one aspect, the characteristic under consideration is a genetic polymorphism or a somatic mutation. In another aspect, the characteristic under consideration is expression level of a gene or a polypeptide. In one aspect, a more desirable clinical outcome is relatively higher likelihood of or relatively better tumor response such as tumor load reduction. In another aspect, a more desirable clinical outcome is relatively longer overall survival. In yet another aspect, a more desirable clinical outcome is relatively longer progression free survival or time to tumor progression. In yet another aspect, a more desirable clinical outcome is relatively longer disease free survival. In further another aspect, a more desirable clinical outcome is relative reduction or delay in tumor recurrence. In another aspect, a more desirable clinical outcome-20-4916-161 1 -5558.1Atty Dkt.: 064189-4890 is relatively decreased metastasis. In another aspect, a more desirable clinical outcome is relatively lower relative risk. In yet another aspect, a more desirable clinical outcome is relatively reduced toxicity or side effects. In some embodiments, more than one clinical outcomes are considered simultaneously. In one such aspect, a patient possessing a characteristic, such as a genotype of a genetic polymorphism, can exhibit more than one more desirable clinical outcomes as compared to patients having the same disease and receiving the same therapy but not possessing the characteristic. As defined herein, the patient is considered suitable for the therapy. In another such aspect, a patient possessing a characteristic can exhibit one or more desirable clinical outcome but simultaneously exhibit one or more less desirable clinical outcome. The clinical outcomes will then be considered collectively, and a decision as to whether the patient is suitable for the therapy will be made accordingly, taking into account the patient’s specific situation and the relevance of the clinical outcomes. In some embodiments, progression free survival or overall survival is weighted more heavily than tumor response in a collective decision making.

[0096] A “tumor response” (TR) refers to a tumor’s response to therapy. A “complete response” (CR) to a therapy refers to the clinical status of a patient with evaluable but non- measurable disease, whose tumor and all evidence of disease have disappeared following administration of the therapy. In this context, a “partial response” (PR) refers to a response that is anything less than a complete response. “Stable disease” (SD) indicates that the patient is stable following the therapy. “Progressive disease” (PD) indicates that the tumor has grown (i.e. become larger) or spread (i.e. metastasized to another tissue or organ) or the overall cancer has gotten worse following the therapy. For example, tumor growth of more than 20 percent since the start of therapy typically indicates progressive disease. “Nonresponse” (NR) to a therapy refers to status of a patient whose tumor or evidence of disease has remained constant or has progressed.

[0097] “ Overall Survival” (OS) refers to the length of time of a cancer patient remaining alive following a cancer therapy.

[0098] “Progression free survival” (PFS) or “Time to Tumor Progression” (TTP) refers to the length of time following a therapy, during which the tumor in a cancer patient does not grow. Progression-free survival includes the amount of time a patient has experienced a complete response, partial response or stable disease.-21-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0099] “Disease free survival” refers to the length of time following a therapy, during which a cancer patient survives with no signs of the cancer or tumor.

[0100] “ Time to Tumor Recurrence (TTR)” refers to the length of time, following a cancer therapy such as surgical resection or chemotherapy, until the tumor has reappeared (come back). The tumor may come back to the same place as the original (primary) tumor or to another place in the body.

[0101] “Relative Risk” (RR), in statistics and mathematical epidemiology, refers to the risk of an event (or of developing a disease) relative to exposure. Relative risk is a ratio of the probability of the event occurring in the exposed group versus a non-exposed group.

[0102] The term “determining” or “identifying” is to associate or affiliate a patient closely to a group or population of patients who likely experience the same or a similar clinical response to a therapy.

[0103] The term “selecting” a patient for a therapy refers to making an indication that the selected patient is suitable for the therapy. Such an indication can be made in writing by, for instance, a handwritten prescription or a computerized report making the corresponding prescription or recommendation.

[0104] When a genetic marker or polymorphism “is used as a basis” for identifying or selecting a patient for a treatment described herein, the genetic marker or polymorphism is measured before and / or during treatment, and the values obtained are used by a clinician in assessing any of the following: (a) probable or likely suitability of an individual to initially receive treatment s); (b) probable or likely unsuitability of an individual to initially receive treatment(s); (c) responsiveness to treatment; (d) probable or likely suitability of an individual to continue to receive treatment(s); (e) probable or likely unsuitability of an individual to continue to receive treatment(s); (f) adjusting dosage; (g) predicting likelihood of clinical benefits; or (h) toxicity. As would be well understood by one in the art, measurement of the genetic marker or polymorphism in a clinical setting is a clear indication that this parameter was used as a basis for initiating, continuing, adjusting and / or ceasing administration of the treatments described herein.-22-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0105] “Having the same cancer” is used when comparing one patient to another or alternatively, one patient population to another patient population. For example, the two patients or patient population will each have or be suffering from colon cancer.

[0106] A “normal cell corresponding to the tumor tissue type” refers to a normal cell from a same tissue type as the tumor tissue. A non-limiting examples is a normal lung cell from a patient having lung tumor, or a normal colon cell from a patient having colon tumor.

[0107] The term “amplification” or “amplify” as used herein means one or more methods known in the art for copying a target nucleic acid, thereby increasing the number of copies of a selected nucleic acid sequence. Amplification can be exponential or linear. A target nucleic acid can be either DNA or RNA. The sequences amplified in this manner form an “amplicon.” While the exemplary methods described hereinafter relate to amplification using the polymerase chain reaction (“PCR”), numerous other methods are known in the art for amplification of nucleic acids (e.g., isothermal methods, rolling circle methods, etc.). The skilled artisan will understand that these other methods can be used either in place of, or together with, PCR methods.

[0108] The term “complement” as used herein means the complementary sequence to a nucleic acid according to standard Watson / Crick base pairing rules. A complement sequence can also be a sequence of RNA complementary to the DNA sequence or its complement sequence, and can also be a cDNA. The term “substantially complementary” as used herein means that two sequences hybridize under stringent hybridization conditions. The skilled artisan will understand that substantially complementary sequences need not hybridize along their entire length. In particular, substantially complementary sequences comprise a contiguous sequence of bases that do not hybridize to a target or marker sequence, positioned 3’ or 5’ to a contiguous sequence of bases that hybridize under stringent hybridization conditions to a target or marker sequence.

[0109] As used herein, the term “hybridize” or “specifically hybridize” refers to a process where two complementary nucleic acid strands anneal to each other under appropriately stringent conditions. Hybridizations are typically conducted with probe-length nucleic acid molecules. Nucleic acid hybridization techniques are well known in the art. Those skilled in the art understand how to estimate and adjust the stringency of hybridization conditions such-23-4916-161 1 -5558.1Atty Dkt.: 064189-4890 that sequences having at least a desired level of complementarity will stably hybridize, while those having lower complementarity will not. For examples of hybridization conditions and parameters, see, e.g., Sambrook, et al., 1989, Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Press, Plainview, N.Y.; Ausubel, F. M. et al. 1994, Current Protocols in Molecular Biology. John Wiley & Sons, Secaucus, N.J.

[0110] “Primer” as used herein refers to an oligonucleotide that is capable of acting as a point of initiation of synthesis when placed under conditions in which primer extension is initiated (e.g., primer extension associated with an application such as PCR). The primer is complementary to a target nucleotide sequence and it hybridizes to a substantially complementary sequence in the target and leads to addition of nucleotides to the 3 ’-end of the primer in the presence of a DNA or RNA polymerase. The 3 ’-nucleotide of the primer should generally be complementary to the target sequence at a corresponding nucleotide position for optimal expression and amplification. An oligonucleotide “primer” can occur naturally, as in a purified restriction digest or can be produced synthetically. The term “primer” as used herein includes all forms of primers that can be synthesized including, peptide nucleic acid primers, locked nucleic acid primers, phosphorothioate modified primers, labeled primers, and the like.[OHl] Primers are typically between about 5 and about 100 nucleotides in length, such as between about 15 and about 60 nucleotides in length, such as between about 20 and about 50 nucleotides in length, such as between about 25 and about 40 nucleotides in length. In some embodiments, primers can be at least 8, at least 12, at least 16, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60 nucleotides in length. An optimal length for a particular primer application can be readily determined in the manner described in H. Erlich, PCR Technology. Principles and Application for DNA Amplification (1989).

[0112] “Probe” as used herein refers to nucleic acid that interacts with a target nucleic acid via hybridization. A probe can be fully complementary to a target nucleic acid sequence or partially complementary. The level of complementarity will depend on many factors based, in general, on the function of the probe. A probe or probes can be used, for example to detect the presence or absence of a mutation in a nucleic acid sequence by virtue of the sequence characteristics of the target. Probes can be labeled or unlabeled, or modified in any of a -24-4916-161 1 -5558.1Atty Dkt.: 064189-4890 number of ways well known in the art. A probe can specifically hybridize to a target nucleic acid.

[0113] Probes can be DNA, RNA or a RNA / DNA hybrid. Probes can be oligonucleotides, artificial chromosomes, fragmented artificial chromosome, genomic nucleic acid, fragmented genomic nucleic acid, RNA, recombinant nucleic acid, fragmented recombinant nucleic acid, peptide nucleic acid (PNA), locked nucleic acid, oligomer of cyclic heterocycles, or conjugates of nucleic acid. Probes can comprise modified nucleobases, modified sugar moieties, and modified internucleotide linkages. A probe can be fully complementary to a target nucleic acid sequence or partially complementary. A probe can be used to detect the presence or absence of a target nucleic acid. Probes are typically at least about 10, 15, 21, 25, 30, 35, 40, 50, 60, 75, 100 nucleotides or more in length.

[0114] “Detecting” as used herein refers to determining the presence of a nucleic acid of interest in a sample or the presence of a protein of interest in a sample. Detection does not require the method to provide 100% sensitivity and / or 100% specificity.

[0115] “Detectable label” as used herein refers to a molecule or a compound or a group of molecules or a group of compounds used to identify a nucleic acid or protein of interest. In some cases, the detectable label can be detected directly. In other cases, the detectable label can be a part of a binding pair, which can then be subsequently detected. Signals from the detectable label can be detected by various means and will depend on the nature of the detectable label. Detectable labels can be isotopes, fluorescent moieties, colored substances, and the like. Examples of means to detect detectable label include but are not limited to spectroscopic, photochemical, biochemical, immunochemical, electromagnetic, radiochemical, or chemical means, such as fluorescence, chemifluorescence, or chemiluminescence, or any other appropriate means.

[0116] “TaqMan® PCR detection system” as used herein refers to a method for real time PCR. In this method, a TaqMan® probe which hybridizes to the nucleic acid segment amplified is included in the PCR reaction mix. The TaqMan® probe comprises a donor and a quencher fluorophore on either end of the probe and in close enough proximity to each other so that the fluorescence of the donor is taken up by the quencher. However, when the probe hybridizes to the amplified segment, the 5 ’-exonuclease activity of the Taq polymerase-25-4916-161 1 -5558.1Atty Dkt.: 064189-4890 cleaves the probe thereby allowing the donor fluorophore to emit fluorescence which can be detected.

[0117] As used herein, the term “sample” or “test sample” refers to any liquid or solid material containing nucleic acids. In suitable embodiments, a test sample is obtained from a biological source (i.e., a “biological sample”), such as cells in culture or a tissue sample from an animal, preferably, a human. In an exemplary embodiment, the sample is a biopsy sample.

[0118] “Target nucleic acid” as used herein refers to segments of a chromosome, a complete gene with or without intergenic sequence, segments or portions a gene with or without intergenic sequence, or sequence of nucleic acids to which probes or primers are designed. Target nucleic acids can include wild type sequences, nucleic acid sequences containing mutations, deletions or duplications, tandem repeat regions, a gene of interest, a region of a gene of interest or any upstream or downstream region thereof. Target nucleic acids can represent alternative sequences or alleles of a particular gene. Target nucleic acids can be derived from genomic DNA, cDNA, or RNA. As used herein, target nucleic acid can be native DNA or a PCR-amplified product.

[0119] As used herein the term “stringency” is used in reference to the conditions of temperature, ionic strength, and the presence of other compounds, under which nucleic acid hybridizations are conducted. With high stringency conditions, nucleic acid base pairing will occur only between nucleic acids that have sufficiently long segments with a high frequency of complementary base sequences. Exemplary hybridization conditions are as follows. High stringency generally refers to conditions that permit hybridization of only those nucleic acid sequences that form stable hybrids in 0.018 M NaCl at 65°C. High stringency conditions can be provided, for example, by hybridization in 50% formamide, 5*Denhardt’s solution, 5*SSC (saline sodium citrate) 0.2% SDS (sodium dodecyl sulfate) at 42°C., followed by washing in 0. l x SSC, and 0.1% SDS at 65°C. Moderate stringency refers to conditions equivalent to hybridization in 50% formamide, 5*Denhardf s solution, 5*SSC, 0.2% SDS at 42°C, followed by washing in 0.2* SSC, 0.2% SDS, at 65°C. Low stringency refers to conditions equivalent to hybridization in 10% formamide, 5*Denhardt’s solution, 6* SSC, 0.2% SDS, followed by washing in l x SSC, 0.2% SDS, at 50°C.-26-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0120] As used herein the term “substantially identical” refers to a polypeptide or nucleic acid exhibiting at least 50%, 75%, 85%, 90%, 95%, or even 99% identity to a reference amino acid or nucleic acid sequence over the region of comparison. For polypeptides, the length of comparison sequences will generally be at least 20, 30, 40, or 50 amino acids or more, or the full length of the polypeptide. For nucleic acids, the length of comparison sequences will generally be at least 10, 15, 20, 25, 30, 40, 50, 75, or 100 nucleotides or more, or the full length of the nucleic acid.

[0121] Modes for Carrying Out the Disclosure

[0122] The disclosure further provides diagnostic, prognostic and therapeutic methods, which are based, at least in part, on determination of the identify of a genotype of interest identified herein.

[0123] For example, information obtained using the diagnostic assays described herein is useful for determining if a subject is suitable for cancer treatment of a given type. Based on the prognostic information, a doctor can recommend a therapeutic protocol, useful for reducing the malignant mass or tumor in the patient or treat cancer in the individual.

[0124] A patient’s likely clinical outcome following a clinical procedure such as a therapy or surgery can be expressed in relative terms. For example, a patient having a particular genotype or expression level can experience relatively longer overall survival than a patient or patients not having the genotype or expression level. The patient having the particular genotype or expression level, alternatively, can be considered as likely to survive. Similarly, a patient having a particular genotype or expression level can experience relatively longer progression free survival, or time to tumor progression, than a patient or patients not having the genotype or expression level. The patient having the particular genotype or expression level, alternatively, can be considered as not likely to suffer tumor progression. Further, a patient having a particular genotype or expression level can experience relatively shorter time to tumor recurrence than a patient or patients not having the genotype or expression level. The patient having the particular genotype or expression level, alternatively, can be considered as not likely to suffer tumor recurrence. Yet in another example, a patient having a particular genotype or expression level can experience relatively more complete response or partial response than a patient or patients not having the genotype or expression level. The-27-4916-161 1 -5558.1Atty Dkt.: 064189-4890 patient having the particular genotype or expression level, alternatively, can be considered as likely to respond. Accordingly, a patient that is likely to survive, or not likely to suffer tumor progression, or not likely to suffer tumor recurrence, or likely to respond following a clinical procedure is considered suitable for the clinical procedure.

[0125] It is to be understood that information obtained using the diagnostic assays described herein can be used alone or in combination with other information, such as, but not limited to, genotypes or expression levels of other genes, clinical chemical parameters, histopathological parameters, or age, gender and weight of the subject. When used alone, the information obtained using the diagnostic assays described herein is useful in determining or identifying the clinical outcome of a treatment, selecting a patient for a treatment, or treating a patient, etc. When used in combination with other information, on the other hand, the information obtained using the diagnostic assays described herein is useful in aiding in the determination or identification of clinical outcome of a treatment, aiding in the selection of a patient for a treatment, or aiding in the treatment of a patient and etc. In a particular aspect, the genotypes or expression levels of one or more genes as disclosed herein are used in a panel of genes, each of which contributes to the final diagnosis, prognosis or treatment.

[0126] In some aspects, the disclosure provides a method for determining whether a prostate cancer patient who is naive to androgen ablation therapy is likely to be responsive to non- hormonal therapy, the method comprising, or consisting essentially of, or consisting of determining whether a prostate tumor sample isolated from the patient contains a an inherent variant polymorphism in an SRD5A gene, wherein the patient is more likely to be responsive to non-hormonal therapy when the patient has the inherent variant polymorphism in the SRD5A gene and is less likely to be responsive to non-hormonal therapy when the patient lacks the inherent variant polymorphism or has wildtype genotype the SRD5A gene. In some aspects, the disclosure provides a method, wherein the SRD5A gene is the SRD5A3 gene, e.g., wherein the SRD5A gene includes the SRD5A3 and the SRD5A1 genes. In a further aspect, the SRD5A gene further includes the SRD5A2 gene.

[0127] In some aspects, the disclosure provides a method, wherein the inherent variant is one or more position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, -28-4916-161 1 -5558.1Atty Dkt.: 064189-4890 rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100).

[0128] In a further aspect, the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0129] In one aspect of the method, the non-hormonal therapy comprises, or consists essentially of, or consists of, administration of one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors. Non-limiting examples include administration of a chemotherapy that comprises, or consisting essentially of, or consists of, includes one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

[0130] In some aspects of the method, the SRD5A genotype is determined by a method comprising, or consisting essentially of, or yet further consisting of one or more of : PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods.

[0131] The patient can be an animal, a mammal or a human patient. In aspects where the patient is a non-human animal, the method provides an animal model for testing effectiveness of new therapies and combination therapies.

[0132] In some aspects, the patient has undergone surgical resection of the prostate tumor before or after the method.

[0133] In some aspects, the diagnostic is useful for the administration of a first-line therapy.

[0134] Therapeutic Methods

[0135] In some aspects, the disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, the method comprising, or consists essentially of or yet further consists of administering an effective amount of a non-hormonal therapy, wherein the patient has an inherent variant genotype for a SRD5A gene.

[0136] In some aspects, the disclosure provides a method, wherein the non-hormonal therapy excludes administration of a combined androgen blockage (CAB) therapy, optionally a-29-4916-161 1 -5558.1Atty Dkt.: 064189-4890 therapy including ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0137] In some aspects, the disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, the method comprising, or consists essentially of or yet further consists of administering an effective amount of a hormonal therapy, wherein the patient lacks an inherent variant genotype for a SRD5A gene.

[0138] In some aspects, the disclosure provides a method, wherein the hormonal therapy is selected from a combined androgen blockage (CAB) therapy, optionally a therapy including ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0139] In some aspects, the disclosure provides a method 1-4, wherein the SRD5A gene is the SRD5A3 gene, or the SRD5A3 and the SRD5A1 genes, or the SRD5A2 gene.

[0140] In some aspects of this disclosure the inherent variant is one or more at a position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100).

[0141] In one aspect, the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0142] In some aspects of the method the non-hormonal therapy includes one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors. In a further aspect, method comprises, or consists essentially of, or yet further consists of administration of a chemotherapy that comprises, or consists essentially of, or yet further consists of administration of one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.-30-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0143] In some aspects, the disclosure provides a method, wherein the inherent variant is one or more at a position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100).

[0144] In some aspects of the method, the SRD5A genotype is determined by a method including one or more of : PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods, optionally using a patient sample selected from a blood sample, a tissue sample such as a tumor sample.

[0145] In another aspect of the method, the patient has undergone surgical resection of the prostate tumor.

[0146] In some aspects, the administration is delivered as a first-line therapy.

[0147] In one embodiment, provided herein are methods for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising, or consisting essentially of, or consisting of administering an effective amount of a non-hormonal therapy to the patient, wherein the patient has an inherent variant genotype for an SRD5A gene. In one aspect, the SRD5A gene is the SRD5A3 gene, optionally at a location identified by one, more or all of rs73236134, rs7674388, rs73236133, rs7663650, rs7664100. In another aspect, the SRD5A gene comprises the SRD5A3 gene, optionally at a location identified by one, more or all of rs73236134, rs7674388, rs73236133, rs7663650, rs7664100) and the SRD5A1 gene, optionally at a location identified by one, more or all of rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930). In yet further aspect, the SRD5A gene further comprises the SRD5A2 gene optionally at a location identified by one, more or all of rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242,-31-4916-161 1 -5558.1Atty Dkt.: 064189-4890 rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063.

[0148] In some aspects, the disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, including administering an effective amount of a non-hormonal therapy, wherein the patient has an inherent variant genotype for a SRD5A gene.

[0149] In some aspects, the disclosure provides a method, wherein the non-hormonal therapy excludes administration of a combined androgen blockage (CAB) therapy, optionally a therapy including ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0150] In some aspects, the disclosure provides a method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, including administering an effective amount of a hormonal therapy, wherein the patient lacks an inherent variant genotype for a SRD5A gene.

[0151] In some aspects, the disclosure provides a method, wherein the hormonal therapy is selected from a combined androgen blockage (CAB) therapy, optionally a therapy including ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0152] In some aspects, the disclosure provides a method 1-4, wherein the SRD5A gene is the SRD5A3 gene.

[0153] In some aspects, the disclosure provides a method, wherein the SRD5A gene includes the SRD5A3 and the SRD5A1 genes.

[0154] In some aspects, the disclosure provides a method, wherein the SRD5A gene further includes the SRD5A2 gene.

[0155] In some aspects, the disclosure provides a method, wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.-32-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0156] In some aspects, the disclosure provides a method, wherein the non-hormonal therapy includes one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibodydrug conjugates, or PARP inhibitors.

[0157] In some aspects, the disclosure provides a method, wherein the chemotherapy includes one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

[0158] In some aspects, the disclosure provides a method, wherein the inherent variant is one or more at a position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100).

[0159] In some aspects, the disclosure provides a method, wherein the SRD5A genotype is determined by a method including one or more of : PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods, optionally using a patient sample selected from a blood sample, a tissue sample such as a tumor sample.

[0160] In some aspects, the disclosure provides a method, wherein the patient has undergone surgical resection of the prostate tumor.

[0161] In some aspects, the disclosure provides a method, wherein the therapy is a first-line therapy.

[0162] Non-limiting examples of non-hormonal therapies comprise one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors. Non-limiting examples of chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

[0163] Men carrying SRD5A gene variants are more likely to experience worse outcomes in response to combined androgen blockade (CAB) therapy, especially when ADT is combined-33-4916-161 1 -5558.1Atty Dkt.: 064189-4890 with a selective CYP 17,20 lyase inhibitor. Thus, this invention provides a method to treat a metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising, or consisting essentially of, or consisting of administering an effective amount of a non-hormonal therapy to the patient, wherein the cancer patient carries an SRD5 A gene variant as identified herein, especially when combined with a selective CYP 17,20 lyase inhibitor.

[0164] In another aspect of this disclosure, the method further comprises, or consists essentially of, or yet further consisting of analyzing a prostate tumor sample isolated from the patient for an inherent variant in a position identified by one or more of 17 in SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100.

[0165] In a further aspect, the SRD5A genotype is determined by a method comprising PCR, whole genome sequencing, or hybridization.

[0166] In another aspect, the patient has undergone surgical resection of the prostate tumor.

[0167] In a yet further aspect, the therapy is a first-line therapy.

[0168] In one aspect, the method is performed by first analyzing an appropriate sample, such as a blood sample or tumor sample from the patient. However, the assay could also be performed on DNA from the patient’s tissues or from the tumor, which would also carry the patient’s inherited variants. Tumor DNA could be obtained either from tumor tissue (fresh, frozen, or paraffin-embedded blocks) or from circulating tumor cells or circulating cell-free DNA isolated from a blood sample. DNA can be isolated from blood or tissues using commercially available kits (eg. Qiagen). Ather isolation and preparation of the sample, the variants can be assayed by any appropriate method, e.g., PCR technology (eg. Taqman), or next generation sequencing technologies such as whole genome sequencing or using microarrays (hybridization technology).-34-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0169] In one aspect, PCR assay kits targeted to specific rs numbers can be purchased from commercial companies (eg. ThermoFisher Assays on Demand (predesigned, available for many, many SNPs) or ThermoFisher Assays by Design (targeted to any specified rs number). PCR assays are carried out using the kits, according to manufacturer’s instructions. Similar assay kits could be purchased for the other 40 rsnumbers as described herein.

[0170] Sequencing can be directed to the entire genome, the entire exome, or to panels of small, targeted DNA segments. The first step is to fragment the DNA and produce a “library” of DNA fragments to be sequenced (eg. using the Illumina DNA prep kit). For targeted sequencing, library preparation additionally involves an enrichment step (using hybridization or multiplex PCR, eg. using Illumina Ampliseq kits) to enrich the sample for the targets of interest. The prepared DNA libraries are then sequenced on a DNA sequencer and the genotypes of interest are extracted from the sequence data output files.

[0171] Microarrays can be used and the advantage of this method is that a predesigned panel of SNPs (eg. the Illumina Global Diversity Array) can indirectly (through linkage disequilibrium) pick up “signals” from the entire genome.

[0172] Prognostic Methods

[0173] Also provided herein are methods for determine whether a prostate cancer patient who is naive to androgen ablation therapy is likely to be responsive to non-hormonal therapy, the method comprising determining whether a prostate tumor sample isolated from the patient contains an inherited variant polymorphism in an SRD5A gene, wherein the patient is more likely to be responsive to non-hormonal therapy when the patient has an inherited variant polymorphism in SRD5A genes and is less likely to be responsive to non-hormonal therapy when the patient has wildtype genotype the SRD5 A gene.

[0174] In one aspect, the SRD5A gene is the SRD5A3 gene, optionally at a location identified by one, more or all of rs73236134, rs7674388, rs73236133, rs7663650, rs7664100. In another aspect, the SRD5A gene comprises the SRD5A3 gene, optionally at a location identified by one, more or all of rs73236134, rs7674388, rs73236133, rs7663650, rs7664100) and the SRD5A1 gene, optionally at a location identified by one, more or all of rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930). In-35-4916-161 1 -5558.1Atty Dkt.: 064189-4890 yet further aspect, the SRD5A gene further comprises the SRD5A2 gene optionally at a location identified by one, more or all of rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063.

[0175] Non-limiting examples of non-hormonal therapies comprise one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors. Non-limiting examples of chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or alimta.

[0176] In a further aspect, the method further comprises administering an effective amount of chemotherapy or radiation therapy to a patient who is identified as likely to be responsive to the non-hormonal therapy. Non-limiting examples of non-hormonal therapies comprise one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors. Non-limiting examples of chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or alimta.

[0177] In a further aspect, the SRD5A genotype is determined by a method comprising PCR, whole genome sequencing or nucleic acid hybridization.

[0178] In a yet further aspect, the patient has undergone surgical resection of the prostate tumor.

[0179] In one aspect, the method is performed by first analyzing an appropriate sample, such as a blood sample or tumor sample from the patient. However, the assay could also be performed on DNA from the patient’s tissues or from the tumor, which would also carry the patient’s inherited variants. Tumor DNA could be obtained either from tumor tissue (fresh, frozen, or paraffin-embedded blocks) or from circulating tumor cells or circulating cell-free DNA isolated from a blood sample. DNA can be isolated from blood or tissues using commercially available kits (eg. Qiagen). Ather isolation and preparation of the sample, the variants can be assayed by any appropriate method, e.g., PCR technology (eg. Taqman), or next generation sequencing technologies such as whole genome sequencing or using microarrays (hybridization technology).-36-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0180] In one aspect, PCR assay kits targeted to specific rs numbers can be purchased from commercial companies (eg. ThermoFisher Assays on Demand (predesigned, available for many, many SNPs) or ThermoFisher Assays by Design (targeted to any specified rs number). PCR assays are carried out using the kits, according to manufacturer’s instructions. Similar assay kits could be purchased for the other 40 rsnumbers as described herein.

[0181] Sequencing can be directed to the entire genome, the entire exome, or to panels of small, targeted DNA segments. The first step is to fragment the DNA and produce a “library” of DNA fragments to be sequenced (eg. using the Illumina DNA prep kit). For targeted sequencing, library preparation additionally involves an enrichment step (using hybridization or multiplex PCR, eg. using Illumina Ampliseq kits) to enrich the sample for the targets of interest. The prepared DNA libraries are then sequenced on a DNA sequencer and the genotypes of interest are extracted from the sequence data output files.

[0182] Microarrays can be used and the advantage of this method is that a predesigned panel of SNPs (eg. the Illumina Global Diversity Array) can indirectly (through linkage disequilibrium) pick up “signals” from the entire genome.

[0183] Experiment No. 1 :

[0184] This experiment establishes the broad significance of the SRD5 A family variants on treatment outcome using a comprehensive genotype panel, application for research or clinical applications.

[0185] 5-alpha steroid reductase isoenzymes are encoded by members of the SRD5A gene family, which includes SRD5A1, SRD5A2, and SRD5A3. In normal prostate and early-stage PCa, SRD5A2 is predominantly expressed, whereas SRD5A1 expression increases with androgen deprivation. SRD5A1 and SRD5A3 are overexpressed in CRPC, and it has been suggested that SRD5A3 may play a role in progression to hormone resistancel3-19. One prior study investigated several SRD5A1 and SRD5A2 gene polymorphisms among 104 Japanese men with metastatic prostate cancer who had undergone primary ADT12. Shiota et al. (Shiota et al. study) reported a higher risk of progression and death among men with the SRD5A2 rs523349 GG genotype vs. other genotypes (GC or CC), though survival was similar for GG vs. CC.13-15 This study was limited by the small sample size, especially the small number (12) of CC homozygous men.-37-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0186] The Shiota et al. study found only one variant in the SRD5A2 gene to be statistically significantly associated with outcome, and this finding was of questionable relevance for several reasons. One is the lack of allelic dose-response. Those men who carried either two copies of the variant allele (genotype CC) or two wildtype alleles (genotype GG) had essentially identical survival curves, with worse survival compared to those men with one wildtype and one variant allele (genotype GC). Thus, the study’s finding lacks biological plausibility. Furthermore, dose-response is one of the criteria used to distinguish causal correlations from chance findings (A. Hill, Proc R Soc Med 58: 295-300; 1965). Lack of dose response plus the small sample size suggests that this is likely to have been a chance finding (false positive).

[0187] In contrast, Applicant’s study, with 1200 men from multiple centers and more diverse genetic backgrounds provides a more complete picture of the genetic factors influencing treatment response.

[0188] Briefly, Applicant studied the inherited variants in the SRD5A gene family and response to hormonal therapy in men with mCSPC enrolled in SWOG S1216, a phase III randomized multicenter clinical trial of ADT with a CYP 17,20 lyase inhibitor (TAK-700) or an androgen receptor inhibitor (bicalutamide)20. Inherited variants (17 in SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100) were assayed using the Illumina Global Diversity Array (GDA). In brief, genomic DNA (300ng) was amplified and then hybridized to an 8-sample array. Primer extension utilizing red- and green-labeled dideoxynucleotides specific for each of the 1.8 million genotypes assayed on the array was performed, followed by Illumina iScan technology imaging. Raw data files (.idat) were generated in both red- and green-fluorescent channels and were used as input for Illumina GenomeStudio software. Genotyping calls were exported using GenomeStudio software for biological interpretation. The Illumina Array Analysis Platform Genotyping Command Line was used to process IDAT (Illumina intensity) files into GTC files. Next, the-38-4916-161 1 -5558.1Atty Dkt.: 064189-4890Bcftoolsvl .16 gtc2vcf plugin was utilized to generate VCF files. VEP were used to annotate and predict the effects of gene variants. For statistical analysis and visualization, Applicant utilized Rv3.6.0 packages: circlize (0.4.6), ComplexHeatmap (1.99.7), dplyr (0.8.0.1), ggplot2 (3.1.1), ggpubr (0.2), maftools (2.0.05), plyr (1.8.4), png (0.1-7), qvalue (2.16.0), reshape2 (1.4.3), stringr (1.4.0), TCGAbiolinks (2.12.6), tidyr (0.8.3), tools (3.6.0).

[0189] Associations with progression free survival (PFS) and overall survival (OS) were analyzed using Kaplan-Meier curves and Cox proportional hazards models. Associations with 7-month PSA response (< 0.2, 0.2-4.0, >4.0 ng / ml) were analyzed using ordinal Chi- Square tests and multinomial logistic regression models. All models were adjusted for treatment arm, extent of disease at study entry (extensive v. minimal), Zubrod performance status, and ADT status at baseline (already initiated or not). Of 364 men with GDA genotype data, 38 (10.4%) carried variant alleles on one or more of the SRD5A genes and 25 (66%) of these men carried variant alleles on all three genes. Variant allele carriers had shorter PFS (HR=3.6; 95% CI 2.5-5.3; p<0.001) and were less likely to experience complete PSA response to treatment (p=0.006). For OS there was a statistically significant interaction between treatment arm and variant allele status (p=0.02). Among those in the TAK-700 arm, median OS was 25.5 months for variant carriers vs. 94.7 months for non-carriers (HR=5.4; 95% CI 2.9-10.1; p<0.001). The association was weaker in the bicalutamide arm: median OS was 39.7 months for variant carriers vs. 86.1 months for non-carriers (HR=1.9; 95% CI 1.1-3.1; p=0.02). Applicant concluded that men carrying SRD5A gene variants may experience worse outcomes in response to hormonal therapy, especially when ADT is combined with a selective CYP 17,20 lyase inhibitor.

[0190] This study makes use of samples and data from the completed SWOG S1216 multicenter randomized clinical trial of androgen deprivation therapy with either a CYP 17,20 lyase inhibitor (TAK-700) or an androgen receptor inhibitor (bicalutamide). The 1200 participants enrolled in this study were men with metastatic hormone sensitive prostate cancer. The median follow-up time was 4.9 years. The study’s primary outcome was overall survival, which did not differ significantly between the two study arms. Secondary outcomes included progression free survival and PSA response at seven months.

[0191] Descriptive statistics: Genotyping was carried out on buffy coat DNA samples from364 participants in the SWOG 1216 study (Table 1). Patient ages ranged from 44 to 88 years,-39-4916-161 1 -5558.1Atty Dkt.: 064189-4890 with a mean and median of 67 years. Most participants (89%) reported their race to be white. Ten percent of participants carried at least one variant allele on at least one of the SRD5A genes. Of the 38 variant carriers, 66% carried variant alleles on all 3 genes, 29% on 2 genes, and 5% carried variant alleles only on the SRD5 A2 gene.

[0192] Table :3 Participants with genotype data (N=364)

[0193] Association of individual variants with disease progression: Each of the 31 SRD5A variants was tested for association with progression free survival using Kaplan-Meier curves and log-rank tests. FIGS. 3 A - 3C show log-rank p values for each variant, arranged according to their position within the gene. All three genes had highly significant associations between variant alleles and disease progression.

[0194] Association of SRD5A carrier status with progression free survival: The relationship between SRD5A carrier status (combined over the 3 genes: any SRD5A variant allele vs. no variant alleles) and progression free survival was further examined using Cox proportional hazards models. All models were adjusted for treatment arm, extent of disease at study entry (extensive vs. minimal), Zubrod performance status, and ADT status at baseline (already initiated or not). Race and age were not statistically significant and were therefore excluded from the models.

[0195] Association of SRD5A carrier status with overall: s a statistically significant interaction between the treatment arm and variant allele status (p=0.02); therefore, models were stratified by treatment arm.4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0196] Association of SRD5A carrier status with PSA response to treatment: Associations of SRD5A carrier status and 7-month PSA response (non-response; partial response; complete response) were analyzed using multinomial logistic regression models (Table 2).

[0197] Table 4: PSA response at 7 months by SRD5A carrier status

[0198] The odds of partial response vs. non-response did not differ significantly in terms of SRD5A carrier status. However, complete response was experienced by 24% of variant allele carriers vs. 55% of non-carriers. After adjusting for treatment arm, extent of disease at study entry (extensive vs. minimal), Zubrod performance status, and ADT status at baseline (already initiated or not), the odds of complete PSA response at 7 months were 72% lower (OR=0.28) for variant allele carriers than for non-carriers (p=0.006).

[0199] Men carrying SRD5A gene variants may experience worse outcomes in response to hormonal therapy (shorter time to disease progression, shorter overall survival, and lesser PSA response to initial therapy), especially when ADT is combined with a selective CYP 17,20 lyase inhibitor.

[0200] Experimental Summary

[0201] Prostate cancer (PCa) is the most prevalent and the second most lethal cancer among American men. For men with newly diagnosed metastatic PCa, hormonal therapy is the mainstay of treatment. While initially effective, the duration of benefit is highly variable, ranging from a few months to several years, and it is difficult to reliably predict which men will benefit and for how long. It is increasingly important to establish the optimal use of hormonal therapy and identify patients most likely to benefit from alternative treatments.

[0202] Thus, in view of these findings, Applicant has developed an assay for distinguishing those with “wildtype” capability to produce 5 alpha-reductase vs. those that have genetically altered capacity to produce 5alpha-reductase. To distinguish “wildtype” from “variant,” Applicant evaluated three genes that encode the 5alpha reductase enzyme. These three genes are part of a gene family but reside on three different chromosomes. There are hundreds of-41-4916-161 1 -5558.1Atty Dkt.: 064189-4890 variants in each of the SRD5A genes. To the best of Applicant’s knowledge, Applicant’s methods capture the vast majority of this variation using only a subset of variants (currently, Applicant’s panel of 41 SNPs across the 3 genes). This is possible because the panel of SNPs can “tag” the other variants that are not genotyped. The tag SNP approach is widely used and is the basis for GWAS (genome wide association studies).

[0203] In contrast, the Shiota et al study did not assay the same locations or the entireSRD5 A2 gene. Their finding focused on a single variant that was hypothesized to produce a version of the SRD5A2 enzyme with slightly altered activity. The Shiota et al. study also did not connect the results of their study with responsiveness to therapy. Indeed, the therapies in use at the time of that study in 2015 are very different from current therapies and did not include combination therapy. Moreover, Applicant found that men with variant SR5A2 status are destined to quickly fail therapy and therefore need initial non-hormonal therapy. Patients with SRD5A polymorphism should receive upfront non-hormonal therapy such as docetaxel because they have a very short response to hormonal therapy. Patients with wildtype SRD5A gene are suitable for treatment with hormonal therapy.

[0204] Experiment No. 2

[0205] Experiment No. 2 extends the clinical relevance of the genomic findings from Experiment No. 1.

[0206] Materials and Methods

[0207] SWOG study population: This study makes use of samples and data from SWOG S 1216, a multi-center randomized Phase III, CTEP-approved trial of androgen deprivation therapy with either a CYP 17,20 lyase inhibitor (TAK-700 / orteronel) or an androgen receptor inhibitor (bicalutamide), which enrolled 1,279 patients with mHSPC. The trial was opened in March 2013 and closed to enrollment in 2018. The median patient age was 68 years, the median PSA at enrollment was 30 ng / mL, and 49% had extensive disease. The median follow-up time was 4.9 years.

[0208] SWOG clinical outcomes / measures of disease progression: The study’s primary outcome was overall survival (OS), which did not differ significantly between the two study arms at study completion.8 OS was defined as the time from random assignment until death resulting from any reason. Additional outcomes examined in this publication include-42-4916-161 1 -5558.1Atty Dkt.: 064189-4890 progression free survival (PFS), which was based on PSA, imaging, and / or clinical progression, and PSA value after seven months of therapy, categorized as Non-Response (>4 ng / ml), Partial Response (0.2 -4 ng / ml), and Complete Response (< 0.2 ng / ml).

[0209] Genotyping and gene selection: From 1279 participants in the SWOG 1216 study, 364 huffy coat DNA samples were randomly selected for genotyping by randomly sampling within quartiles of PFS. Variants were assayed in the USC Norris Molecular Genomics Core Shared Resource using the Illumina Global Diversity Array (GDA) with the BeadLab system automated protocol. Eighteen genes (LHRH, LHRH-R, LH, LHR, StAR, CYP11 Al, CYP17A1, HSD3B2, HSD17B2, HSD17B3, SRD5A1, SRD5A2, SRD5A3, RDH5, AKR1C1, AKR1C2, AKR1C3, and CyB5), selected due to their role in androgen synthesis and signaling, were screened for associations with PFS using log-rank tests. Of these genes, only SRD5A1, SRD5A2, and SRD5A3 showed strong, highly significant associations with PFS across multiple SNPs and thus were included in these subsequent analyses. Genotypes were available for 41 variants in these genes: 17 in SRD5A1 (rs477930, rs8192139, rs535981, rs562461, rs248805, rs3822430, rs8192206, rs4702381, rs8192249, rs3297, rsl042150, rsl6877779, rsl2716180, rs79421055, rs28594474, rsl651071, rs2434525); 19 in SRD5A2 (rs28383087, rs28383086, rs28383085, rs9332975, rs28383083, rsl92604242, rs28383082, rsl2470143, rs2366063, rs6727380, rs551741548, rs72794634, rs781063043, rs2268796, rs2300697, rsl988909, rs74403699, rs632148, rs559555); and 5 in SRD5A3 (rs73236134, rs73236133, rs7674388, rs7664100, rs7663650).

[0210] Statistics: Due to strong correlations among the 41 variants, SRD5A variant allele carrier status was defined as a dichotomous variable: carrying no variant alleles on all three SRD5A genes vs. carrying at least one variant allele on at least one of the three genes. Associations of SRD5A carrier status with PFS and OS were analyzed using Kaplan-Meier curves and Cox proportional hazards models. Associations with 7-month PSA response (< 0.2, 0.2-4.0, >4.0 ng / ml) were analyzed using ordinal Chi-Square tests and multinomial logistic regression models. All models were adjusted for treatment arm, extent of disease at study entry (extensive v. minimal), Zubrod performance status, and ADT status at baseline (already initiated or not). Race and age were excluded from the models due to lack of statistical significance. Interactions between treatment arm and SRD5 A carrier status were evaluated using the Wald test to assess statistical significance.-43-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0211] Data Availability: The data generated in this study will be made publicly available in ClinVar.

[0212] Results

[0213] Descriptive statistics: Patients were similar to those in the overall cohort. Ages ranged from 44 to 88 years, with a mean and median of 67 years. Most participants (89%) reported their race to be white. Fifty-one percent had extensive disease. Median follow-up time was 6 years. Ninety percent (90%) of participants carried no variant alleles for all three SRD5A genes. Ten percent (10%) of participants carried at least one variant allele on at least one of the SRD5A genes. Of the 38 variant carriers, 66% carried variant alleles on all 3 genes, 29% on 2 genes, and 5% carried variant alleles only on the SRD5 A2 gene. All variant allele carriers tested positive for 4 or more of the 41 variants assayed (17 in SRD5A1, 19 in SRD5A2, and 5 in SRD5A3). The median number of variants was 15, and two men carried 22 of the 41 variant alleles.

[0214] Association of individual variants with disease progression: All three SRD5 A genes showed highly significant associations between individual variant alleles and PFS. Using log-rank tests, 15 of 17 SRD5A1 variants, 12 of 19 SRD5A2 variants, and 4 of 5 SRD5A3 variants were significant with p<0.01. P values <10-5 were observed for 9 of 17 SRD5A1 variants, 10 of 19 SRD5A2 variants, and 2 of 5 SRD5A3 variants.

[0215] Association of SRD5A carrier status with PFS and OS: For PFS (FIG. 6A), variant allele carriers had shorter PFS than non-carriers in multivariable adjusted Cox proportional hazard models (HR=3.6; 95% CI 2.5-5.3; p<0.001). Median PFS was 9.9 months for variant carriers vs. 41.0 months for non-carriers. For overall survival (FIG. 6B), there was a statistically significant interaction between the treatment arm and variant allele status (p=0.02); therefore, models were stratified by treatment arm. Among those in the TAK-700 arm (ADT plus a CYP 17,20 lyase inhibitor), median overall survival was 25.5 months for variant carriers vs. 94.7 months for non-carriers (HR=5.4; 95% CI 2.9-10.1; p<0.001). In the Bicalutamide arm (ADT plus an androgen receptor inhibitor), median overall survival was 39.7 months for variant carriers vs. 86.1 months for non-carriers (HR=1.9; 95% CI 1.1 -3.1; p=0.02).-44-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0216] Association of SRD5A carrier status with 7-month PSA response to treatment: The presence of a variant allele was significantly associated with not achieving a complete PSA response at 7 months (p=0.006). Complete response (PSA < 0.2 ng / ml) was experienced by 24% of variant allele carriers vs. 55% of non-carriers. The odds of complete PSA response at 7 months were 72% lower (OR=0.28, 95% Confidence interval (0.11, 0.69)) for variant allele carriers than for non-carriers. The odds of partial response vs. non-response did not differ significantly with respect to SRD5A carrier status (OR=1.19, 95% Confidence interval (0.53, 2-67)).

[0217] Discussion

[0218] For men with mHSPC, ADT in combination with AR pathway inhibitors (ARPIs) plus / minus 6 cycles of docetaxel is the mainstay of treatment9. While initially effective, the duration of benefit from the standard systemic therapy is highly variable, ranging from a few months to several years, and it is difficult to reliably predict at baseline the duration of benefit with the currently approved therapies.9-11

[0219] A few biomarkers have been investigated as early predictors of response to hormonal therapy. Circulating tumor cells (CTCs) have shown promise as prognostic markers. Applicant’s group recently documented that baseline CTC count, after adjusting for disease burden, is highly prognostic of 7-month PSA and 2-year PFS in mHSPC treated with ADT combined with either bicalutamide or the CYP17 inhibitor, TAK70012. Inherited genetic variants are another potential early predictor of treatment response. Under castrate testosterone levels produced by hormonal therapies, minor variations in gene expression may influence DHT availability in the prostate, thereby impacting treatment response. This paradigm is exemplified by the HSD3B1 gene, which possesses adrenal-restrictive and adrenal-permissive alleles, encoding 3BHSD1 enzymes that differ in stability. The adrenal- permissive allele results in increased production of DHT13 and has been associated with worse PFS and OS in response to ADT in several cohorts.14

[0220] One prior study investigated several SRD5A1 and SRD5A2 gene polymorphisms among 104 Japanese men with metastatic prostate cancer who had undergone primary ADT15. Shi ota et al reported a higher risk of progression and death among men with the SRD5A2 rs523349 GG genotype vs. other genotypes (GC or CC), though survival was-45-4916-161 1 -5558.1Atty Dkt.: 064189-4890 similar for GG vs. CC. This study was limited by the small sample size, especially the small number (12) of CC homozygous men.

[0221] Applicant’s study, in contrast, is larger, includes a more genetically diverse population, and utilizes a prospective randomized clinical trial of newer AR signaling pathway inhibitors used alongside ADT. Applicant’s study examined all three SRD5A genes using SNPs from the Illumina Global Diversity Array, which was designed to capture the vast majority of all exome variation. Using a panel of 41 SRD5A SNPs, Applicant determined that approximately 10% of men in Applicant’s study population carry multiple SRD5A gene variants and these men had a high prevalence of early failure of hormone therapy.

[0222] The mechanism through which hormonal therapy response may be diminished in patients with SRD5 A variants remains unclear and warrants further investigation. Evidence attributing functional effects to specific SNPs is sparse and mostly correlational. In one study of 526 men with prostate cancer and not receiving hormone treatment, sex-steroid levels were measured by mass spectrometry in plasma and prostatic tissue. Of 7 SNPs that were assayed (5 in SRD5A2 and 2 in SRD5A1), five were associated with alterations in profiles of DHT and its metabolites, predominantly with circulating levels of inactive glucuronides. One of these SNPs, SRD5A2 rsl2470143, which was associated with high enzyme activityl6, was genotyped in Applicant’s study. Of the SRD5A variant carriers in Applicant’s study, 74% carried the rsl2470143 variant allele. When repeating Applicant’s analyses using rsl2470143 instead of overall SRD5A carrier status, Applicant’s results were similar but slightly attenuated (eg. the hazard ratio for PFS decreased from 3.63 to 3.18), indicating that the panel of SRD5A variants may better capture risk of treatment failure than does the rs 12470143 SNP alone.

[0223] Applicant proposes that the SNP panel genotyped in this study may be a biomarker for inherited 5-alpha steroid reductase functional differences that become clinically relevant under castrate testosterone levels. It is not known whether these putative functional variants are directly assayed by Applicant’s genotyped panel or are unmeasured variants, tagged through linkage disequilibrium by a single SNP or set of SNPs in Applicant’s panel. In either case, a blood-based assay derived from Applicant’s observations would offer a straightforward, cost-effective diagnostic tool. This assay could function as an independent predictor or be integrated within a nomogram alongside other biological markers.-46-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0224] Identification of patients with mHSPC who are unlikely to derive significant benefit from ADT-based hormonal blockade therapies is of critical clinical relevance, as these individuals may require prompt initiation of alternative therapeutic agents such as docetaxel or a preferential enrollment in a clinical trial. Conversely, patients not harboring SRD5A risk alleles may respond favorably to ADT with androgen receptor pathway inhibitors, thus potentially mitigating the need for additional therapies. Given a significantly longer overall survival, these patients may be preferred for ongoing de-escalation trials in the mHSPC setting.

[0225] After adjusting for biopsy sampling density, the 5a-reductase inhibitor finasteride significantly reduced the risk of prostate cancer relative to placebo across multiple Gleason scores in the Prostate Cancer Prevention Trial (PCPT), including the most frequently detected intermediate- and high-grade cancers (Gleason scores 6 and 7)17. Whether the addition of a 5a-reductase inhibitor to standard hormonal therapy in mHSPC patients who are carriers of SRD5 A variants could improve treatment response remains unknown and warrants investigation in future clinical trials.

[0226] In conclusion, Applicant’s study demonstrates that patients with mHSPC who carry polymorphisms in the SRD5A gene family may exhibit limited response to ADT with either a CYP17 inhibitor or a first-generation androgen receptor blocker. These data have implications for patient counseling and clinical trial selection. Applicant’s findings support a personalized therapeutic approach for this patient population.

[0227] Experiment No. 3

[0228] Single Nucleotide Polymorphism (SNP) Genotyping Assays

[0229] This experiment demonstrates that the effect seen in Experiment Nos. 1 and 2 can be captured by a single, easily assayed variant (e.g., SDR5A2 rs632148), supporting streamlined clinical biomarker use.

[0230] Materials and Methods

[0231] Genomic DNA was extracted from the blood samples of SWOG clinical trial participants. The SRD5A2 rs632148 polymorphism was genotyped using a pre-designed TaqMan® SNP assay kit (Cat. No. 4351374, Thermo Fisher Scientific, California, USA)-47-4916-161 1 -5558.1Atty Dkt.: 064189-4890 according to the manufacturer’s instructions on an ABI Real-Time PCR system (Applied Biosystems, California, USA). For each reaction, TaqMan® Universal Master Mix (Thermo Fisher Scientific, Delaware, USA) and genomic DNA were dispensed into a 384-well plate. The plates were sealed, gently mixed, and briefly centrifuged.

[0232] PCR amplification was performed using the following cycling conditions: 95°C for 10 minutes, followed by 40 cycles of 92°C for 15 seconds and 60°C for 1 minute. DNA samples with known genotypes were included as positive controls in each run, while water- only no-template controls were used to detect possible contamination. Allele discrimination was conducted using ABI 7500 software (Applied Biosystems) in accordance with the manufacturer’s guidelines.

[0233] Patient samples are sourced from the S 1216 clinical trial. Because of the strong correlation of this single SNP with the other 40 SNPs in SRD5A genes which were previously measured in 364 men using the Illumina Global Diversity Array (Experiment No. 2), SRD5A polymorphism can predict the duration of response to hormonal therapy in metastatic prostate cancer patients.

[0234] Clauses

[0235] Clause 1. A method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising administering an effective amount of a non-hormonal therapy, wherein the patient has an inherent variant genotype for a SRD5A gene.

[0236] Clause 2. The method of clause 1, wherein the non-hormonal therapy excludes administration of a combined androgen blockage (CAB) therapy, optionally a therapy comprising ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0237] Clause 3. A method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising administering an effective amount of a hormonal therapy, wherein the patient lacks an inherent variant genotype for a SRD5A gene.-48-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0238] Clause 4. The method of clause 3, wherein the hormonal therapy is selected from a combined androgen blockage (CAB) therapy, optionally a therapy comprising ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

[0239] Clause 5. The method of any of any one of clauses 1-4, wherein the SRD5A gene is the SRD5A3 gene.

[0240] Clause 6. The method of any one of clauses 1-5, wherein the SRD5A gene comprises the SRD5A3 and the SRD5A1 genes.

[0241] Clause 7. The method of clause 5 or 6, wherein the SRD5A gene further comprises the SRD5A2 gene.

[0242] Clause 8. The method of any one of clauses 1-4, wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0243] Clause 9. The method of any one of clauses 1, 2 or 3-8, wherein the non-hormonal therapy comprises one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors.

[0244] Clause 10. The method of any one of clauses 1-9, wherein the chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

[0245] Clause 11. The method of any one of clauses 1-7, 9 or 10, wherein the inherent variant is one or more at a position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100, optionally wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0246] Clause 12. The method of any one of clauses 1-11, wherein the SRD5A genotype is determined by a method comprising one or more of : PCR, hybridization, whole genome-49-4916-161 1 -5558.1Atty Dkt.: 064189-4890 sequencing, or TaqMan® SNP assay following manufacturers methods, optionally using a patient sample selected from a blood sample, a tissue sample such as a tumor sample.

[0247] Clause 13. The method of any one of clauses 1-12, wherein the patient has undergone surgical resection of the prostate tumor.

[0248] Clause 14. The method of any one of clauses 1-13, wherein the therapy is a first-line therapy.

[0249] Clause 15. A method for determine whether a prostate cancer patient who is naive to androgen ablation therapy is likely to be responsive to non-hormonal therapy, the method comprising determining whether a prostate tumor sample isolated from the patient contains a an inherent variant polymorphism in an SRD5A gene, wherein the patient is more likely to be responsive to non-hormonal therapy when the patient has the inherent variant polymorphism in the SRD5A gene and is less likely to be responsive to non-hormonal therapy when the patient lacks the inherent variant polymorphism or has wildtype genotype the SRD5A gene.

[0250] Clause 16. The method of clause 15, wherein the SRD5A gene is the SRD5A3 gene.

[0251] Clause 17. The method of clause 15 or 16, wherein the SRD5A gene comprises the SRD5A3 and the SRD5A1 genes.

[0252] Clause 18. The method of clause 15 or 16, wherein the SRD5A gene further comprises the SRD5A2 gene.

[0253] Clause 19. The method of any one of clauses 15-18, wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0254] Clause 20. The method of any one of clauses 15-19, wherein the non-hormonal therapy comprises one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors.

[0255] Clause 21. The method of clause 20, wherein the chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

[0256] Clause 22. The method of any one of clauses 15-18, 21 or 22, wherein the inherent variant is one or more position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474,-50-4916-161 1 -5558.1Atty Dkt.: 064189-4890 rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100, optionally wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

[0257] Clause 23. The method of any one of clauses 15-22, wherein the SRD5A genotype is determined by a method comprising one or more of : PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods.

[0258] Clause 24. The method of any one of clauses 15-23, wherein the patient has undergone surgical resection of the prostate tumor.

[0259] Clause 25. The method of any one of clauses 15-24, wherein the therapy is first-line therapy.

[0260] Equivalents

[0261] Thus, it should be understood that although the present disclosure has been specifically disclosed by preferred embodiments and optional features, modification, improvement and variation of the disclosure embodied therein herein disclosed can be resorted to by those skilled in the art, and that such modifications, improvements and variations are considered to be within the scope of this disclosure. The materials, methods, and examples provided here are representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the disclosure.

[0262] The disclosure has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the disclosure. This includes the generic description of the disclosure with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0263] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.-51-4916-161 1 -5558.1Atty Dkt.: 064189-4890

[0264] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference individually. Severl references are identified by an Arabic number, and the full bibliographic citation or these references are provided below. In case of conflict, the present specification, including definitions, will control.

[0265] Other aspects are set forth within the following claims.4916-161 1 -5558.1Atty Dkt.: 064189-4890ReferencesExperiment No. 11. Barata PC, Sartor AO. Metastatic castration-sensitive prostate cancer: Abiraterone, docetaxel, or. Cancer. 2019; 125(11): 1777-1788.2. Ross RW, Xie W, Regan MM, et al. Efficacy of androgen deprivation therapy (ADT) in patients with advanced prostate cancer: association between Gleason score, prostatespecific antigen level, and prior ADT exposure with duration of ADT effect. Cancer. 2008;112(6): 1247-1253.3. Scott WW, Menon M, Walsh PC. Hormonal Therapy of Prostatic Cancer. Cancer. 1980;45 Suppl 7: 1929-1936.4. Duffy, M.J. Biomarkers for prostate cancer: Prostate-specific antigen and beyond. Clin. Chem. Lab. Med. (CCLM) 2020, 58, 326-339.5. Hussain, M.; Tangen, C.M.; Higano, C.; Schelhammer, P.F.; Faulkner, J.; Crawford, E.D.; Wilding, G.; Akdas, A.; Small, E.J.; Donnelly, B.; et al. Absolute prostate-specific antigen value after androgen deprivation is a strong independent predictor of survival in new metastatic prostate cancer: Data from Southwest Oncology Group Trial 9346 (INT-0162). J. Clin. Oncol. 2006, 24,3984-3990.6. Goldkorn A, Tangen C, Piets M, Morrison GJ, Cunha A, Xu T, Pinski JK, Ingles SA, Triche T, Harzstark AL, Kohli M, MacVicar GR, Vaena DA, Crispino AW, McConkey DJ, Lara PN Jr, Hussain MHA, Quinn DI, Vogelzang NJ, Thompson IM Jr, Agarwal N. Baseline Circulating Tumor Cell Count as a Prognostic Marker of PSA Response and Disease Progression in Metastatic Castrate-Sensitive Prostate Cancer (SWOG S1216). Clin Cancer Res. 2021 Apr 1;27(7): 1967-1973. doi: 10.1158 / 1078-0432.CCR-20-3587. Epub 2021 Jan 26. PMID: 33500355; PMCID: PMC8026618.7. Hashimoto, K.; Tabata, H.; Shindo, T.; Tanaka, T.; Hashimoto, J.; Inoue, R.; Muranaka, T.; Hotta, H.; Yanase, M.; Kunishima, Y.; et al. Serum testosterone level is a useful biomarker for determining the optimal treatment for castration-resistant prostate cancer. Urol. Oncol. 2019, 37, 485-491.-53-4916-161 1 -5558.1Atty Dkt.: 064189-48908. Scher, H I.; Lu, D.; Schreiber, N.A.; Louw, J.; Graf, R.P.; Vargas, H.A.; Johnson, A.; Jendrisak, Bambury, R.; Danila, D.; et al. Association of AR-V7 on Circulating Tumor Cells as a Treatment-Specific Biomarker with Outcomes and Survival in Castration-Resistant Prostate Cancer. JAMA Oncol. 2016, 2, 1441-14499. Antonarakis, E.S.; Lu, C.; Wang, H.; Luber, B.; Nakazawa, M.; Roeser, J.C.; Chen, Y.; Mohammad, T.A.; Chen, Y.; Fedor, H.L.; et al. AR-V7 and Resistance to Enzalutamide and Abiraterone in Prostate Cancer. N. Engl. J. Med. 2014, 371, 1028-1038.10. Sabharwal, N.; Sharifi, N. HSD3B1 genotypes conferring adrenal-restrictive and adrenal-permissive phenotypes in prostate cancer and beyond. Endocrinology 2019, 160, 2180-2188. Cancers 2021, 13, 5723 17 of 1911. Hearn, J.W.D.; Sweeney, C.J.; Almassi, N.; Reichard, C.A.; Reddy, C.A.; Li, H.; Hobbs, B.; Jarrard, D.F.; Chen, Y.-H.; Dreicer, R.; et al. HSD3B1 Genotype and Clinical Outcomes in Metastatic Castration-Sensitive Prostate Cancer. JAMA Oncol. 202012. Shiota M, Fujimoto N, Yokomizo A, Takeuchi A, Itsumi M, Inokuchi J, Tatsugami K, Uchiumi T, Naito S. SRD5A gene polymorphism in Japanese men predicts prognosis of metastatic prostate cancer with androgen-deprivation therapy. Eur J Cancer. 2015 Sep;51(14): 1962-9. doi: 10.1016 / j.ejca.2015.06.122. Epub 2015 Jul l l. PMID: 26169017.13. Salam MT, Ursin G, Skinner EC, Dessissa T, Reichardt JK. Associations between polymorphisms in the steroid 5-alpha reductase type II (SRD5A2) gene and benign prostatic hyperplasia and prostate cancer. Urol Oncol. 2005 Jul-Aug;23(4):246-53. doi:10.1016 / j.urol one.2004.12.014. PMID: 16018939.14. Henriquez-Hernandez LA, Valenciano A, Foro-Amalot P, Alvarez-Cubero MJ, Cozar JM, Suarez-Novo JF, Castells-Esteve M, Fernandez-Gonzalo P, De-Paula-Carranza B, Ferrer M, Guedea F, Sancho-Pardo G, Craven-Bartle J, Ortiz-Gordillo MJ, Cabrera-Roldan P, Rodriguez-Melcon JI, Herrera-Ramos E, Rodriguez-Gallego C, Lara PC. Genetic variations in genes involved in testosterone metabolism are associated with prostate cancer progression: A Spanish multicenter study. Urol Oncol. 2015 Jul;33(7):331.el-7. doi:10.1016 / j.urol one.2015.04.003. Epub 2015 May 7. PMID: 25960412.-54-4916-161 1 -5558.1Atty Dkt.: 064189-489015. Bamodu OA, Tzou KY, Lin CD, Hu SW, Wang YH, Wu WL, Chen KC, Wu CC. Differential but Concerted Expression of HSD17B2, HSD17B3, SHBG and SRD5A1 Testosterone Tetrad Modulate Therapy Response and Susceptibility to Disease Relapse in Patients with Prostate Cancer. Cancers (Basel). 2021 Jul 12; 13(14):3478. doi: 10.3390 / cancersl3143478. PMID: 34298692; PMCID: PMC8303483.16. Fang C, Guo ZQ, Chen XY, Liu TZ, Zeng XT, Wang XH. Relationship between SRD5A2 rs9282858 polymorphism and the susceptibility of prostate cancer: A meta-analysis based on 20 publications. Medicine (Baltimore). 2017 May;96(19):e6791. doi: 10.1097 / MD.0000000000006791. PMID: 28489754; PMCID: PMC5428588.17. Cicek MS, Conti DV, Curran A, Neville PJ, Paris PL, Casey G, Witte JS. Association of prostate cancer risk and aggressiveness to androgen pathway genes: SRD5A2, CYP17, and the AR. Prostate. 2004 Apr l;59(l):69-76. doi: 10.1002 / pros.10358. PMID: 14991867.18. Sarma AV, Dunn RL, Lange LA, Ray A, Wang Y, Lange EM, Cooney KA. Genetic polymorphisms in CYP17, CYP3A4, CYP19A1, SRD5A2, IGF-1, and IGFBP-3 and prostate cancer risk in African- American men: the Flint Men’s Health Study. Prostate. 2008 Feb 15;68(3):296-305. doi: 10.1002 / pros.20696. PMID: 18163429; PMCID: PMC2712831.19. Scariano JK, Treat E, Alba F, Nelson H, Ness SA, Smith AY. The SRD5A2 V89L polymorphism is associated with severity of disease in men with early onset prostate cancer. Prostate. 2008 Dec 1;68(16): 1798-805. doi: 10.1002 / pros.20842. PMID: 18780294.20. Agarwal N, Tangen CM, Hussain MHA, Gupta S, Piets M, Lara PN, Harzstark AL, Przemyslaaw W Twardowski W, Paller CJ, Zylla D, Zibelman MR, Levine E, Roth BJ, Goldkorn A, Vaena DA, Kohli M, Crispino T, Vogelzang NJ, Thompson IM, Quinn DI. Orteronel for metastatic hormone-sensitive prostate cancer: A multicenter, ranndomized, open-label Phse III trial (SWOG-1216). J. Clin. Oncol 40:3301-3309; 202221. Hussain M, Goldman B, Tangen C, Higano CS, Petrylak DP, Wilding G, Akdas AM, Small EJ, Donnelly BJ, Sundram SK, Burch PA, Dipaola RS, Crawford ED. Prostate-specific antigen progression predicts overall survival in patients with metastatic prostate cancer: data from Southwest Oncology Group Trials 9346 (Intergroup Study 0162) and 9916. J Clin Oncol. 2009 May 20;27(15):2450-6. doi: 10.1200 / JC0.2008.19.9810. Epub 2009 Apr 20. PMID: 19380444; PMCID: PMC2684851.-55-4916-161 1 -5558.1Atty Dkt.: 064189-489022. Uno, H., Cai, T., Pencina, M. J., D’Agostino, R. B., & Wei, L. J. “On the C-stat sties for evaluating overall adequacy of risk prediction procedures with censored survival data”. Statistics in Medicine 30(10): 1105—1117; 2011.23. J. Lambert and S. Chevret, “Summary measure of discri ination in survival models based on cumulative / dynamic time-dependent ROC curves,” Statistical Methods in Medical Research, 2014.24. Freedman LS. Tables of the number of patients required in clinical trials using the logrank test. Statistics in Medicine 1 : 121-129. https: / / doi.org / 10.1002 / sim.4780010204.Experiment No. 21. Miller WL, Auchus RJ. The molecular biology, biochemistry and physiology of human steroidogenesis and its disorders. Endocr Rev 2011;32:81-151.2. Auchus RJ. The backdoor pathway to dihydrotestosterone. Trends Endocrinol Metab 2004;15:432-438.3. Ghayee HK, Auchus RJ. Clinical implications of androgen synthesis via 5a-reduced precursors. Endocr Dev 2008;13:55-66.4. Penning TM, Detlefsen A. Intracrinology-revisited and prostate cancer. J Steroid Biochem Mol Biol 2020; 196: 105499.5. Hamid ARAH, Putra HWK, Sari NP, Diana P, Sesari SS, Novita E, Gultom FL, Saraswati M, Tanurahardja B, Asmarinah, Umbas R, Mochtar CA. Early upregulation of AR and steroidogenesis enzyme expression after 3 months of androgen-deprivation therapy. BMC Urol 2020; 20:71.6. Chang K-H, Li R, Papari-Zareei M, Shariffi N. Dihydrotestosterone synthesis bypasses testosterone to drive castration-resistant prostate cancer. PNAS 2011; 108:13728-137337. Uemura M, Tamura K, Chung S, Honma S, Okuyama A, Nakamura Y, Nakagawa H. Novel 5 alpha-steroid reductase (SRD5A3, type-3) is overexpressed in hormone-refractory prostate cancer. Caner Sci 2008; 99:81-86.8. Parikh M, Tangen C, Hussain MHA, Gupta S, Callis S, Jo Y, Harzstark A, Paller CJ, George S, Zibelman MR, Cheng HH, Maughan BL, Zhang J, Pachynski RK, Bryce AH, Lin-56-4916-161 1 -5558.1Atty Dkt.: 064189-4890DW, Quinn DI, Lerner SP, Thompson IM, Dorff TB, Lara PN, Agarwal N. Three- and Seven-month Prostate-specific Antigen Levels as Prognostic Markers for Overall Survival in Metastatic Hormone-sensitive Prostate Cancer: Results from SWOG S1216, a Phase 3 Randomized Trial of Androgen Deprivation Plus Orteronel or Bicalutamide. Eur Urol Oncol. 2024; 7: 1097-1104.9. Barata PC, Sartor AO. Metastatic castration-sensitive prostate cancer: Abiraterone, docetaxel, or. Cancer. 2019; 125(11): 1777-1788.10. Ross RW, Xie W, Regan MM, et al. Efficacy of androgen deprivation therapy (ADT) in patients with advanced prostate cancer: association between Gleason score, prostatespecific antigen level, and prior ADT exposure with duration of ADT effect. Cancer. 2008;112(6): 1247-1253.11. Scott WW, Menon M, Walsh PC. Hormonal Therapy of Prostatic Cancer. Cancer. 1980;45 Suppl 7: 1929-1936.12. Goldkorn A, Tangen C, Piets M, Morrison GJ, Cunha A, Xu T, Pinski JK, Ingles SA, Triche T, Harzstark AL, Kohli M, MacVicar GR, Vaena DA, Crispino AW, McConkey DJ, Lara PN Jr, Hussain MHA, Quinn DI, Vogelzang NJ, Thompson IM Jr, Agarwal N. Baseline Circulating Tumor Cell Count as a Prognostic Marker of PSA Response and Disease Progression in Metastatic Castrate-Sensitive Prostate Cancer (SWOG S1216). Clin Cancer Res. 2021 Apr 1;27(7): 1967-1973.13. Sabharwal, N.; Sharifi, N. HSD3B1 genotypes conferring adrenal-restrictive and adrenal-permissive phenotypes in prostate cancer and beyond. Endocrinology 2019, 160, 2180-2188. Cancers 2021, 13, 5723 17 of 1914. Sharifi N, Diaz R, Lin H-M, Roberts E, Horvath LG, Martin A, Stockler MR, Yip S, Subhash VV, Portman N, Davis ID, Sweeney CJ. Survival of men with metastatic hormonesensitive prostate cancer and adrenal-permissive HSD3B1 inheritance. J Clin Invest. 2024; 134(18): 183583.15. Shi ota M, Fujimoto N, Yokomizo A, Takeuchi A, Itsumi M, Inokuchi J, Tatsugami K, Uchiumi T, Naito S. SRD5A gene polymorphism in Japanese men predicts prognosis of metastatic prostate cancer with androgen-deprivation therapy. Eur J Cancer. 2015-57-4916-161 1 -5558.1Atty Dkt.: 064189-4890Sep;51(14): 1962-9. doi: 10.1016 / j.ejca.2015.06.122. Epub 2015 Jul l l. PMID: 26169017.16. Levesque E, Laverdiere I, Lacombe L, Caron P, Rouleau M, Turcotte V, Tetu B, Fradet Y, Guillemette C. Importance of 5a-reductase gene polymorphisms on circulating and intraprostatic androgens in prostate cancer. Clin Cancer Res. 2014 Feb l;20(3):576-84. doi: 10.1158 / 1078-0432. CCR-13-1100. Epub 2013 Nov 25. PMID: 24277450.17. Thompson IM, Paul er Ankerst D, Chi C, Goodman PJ, Tangen CM, Lippman SM, Lucia MS, Pames HL, Coltman CA Jr. Prediction of prostate cancer for patients receiving finasteride: results from the Prostate Cancer Prevention Trial. J Clin Oncol. 2007 Jul 20;25(21):3076-81. doi: 10.1200 / JC0.2006.07.6836. PMID: 17634486.18. Agarwal N, Tangen CM, Hussain MHA, Gupta S, Piets M, Lara PN, Harzstark AL, Twardowski PW, Paller CJ, Zylla D, Zibelman MR, Levine E, Roth BJ, Goldkom A, Vaena DA, Kohli M, Crispino T, Vogelzang NJ, Thompson IM Jr, Quinn DI. Orteronel for Metastatic Hormone-Sensitive Prostate Cancer: A Multicenter, Randomized, Open-Label Phase III Trial (SWOG-1216). J Clin Oncol. 2022 Oct l;40(28):3301-3309. doi:10.1200 / JCO.21.02517. Epub 2022 Apr 21. PMID: 35446628; PMCID: PMC9553390.-58-4916-161 1 -5558.1

Claims

Atty Dkt.: 064189-4890WHAT IS CLAIMED IS:

1. A method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising administering an effective amount of a non-hormonal therapy, wherein the patient has an inherent variant genotype for a SRD5A gene.

2. The method of claim 1, wherein the non-hormonal therapy excludes administration of a combined androgen blockage (CAB) therapy, optionally a therapy comprising ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

3. A method for treating a patient with metastatic castration sensitive prostate cancer who is naive to testosterone suppressive therapy, comprising administering an effective amount of a hormonal therapy, wherein the patient lacks an inherent variant genotype for a SRD5A gene.

4. The method of claim 3, wherein the hormonal therapy is selected from a combined androgen blockage (CAB) therapy, optionally a therapy comprising ADT and a selected CYP 17,20 lyase inhibitor, or an androgen receptor signaling inhibitor, optionally selected from enzalutamide, apalutamide, darolutamide, and abiraterone.

5. The method of any of any one of claims 1-4, wherein the SRD5A gene is the SRD5A3 gene.

6. The method of any one of claims 1-5, wherein the SRD5A gene comprises the SRD5A3 and the SRD5A1 genes.

7. The method of claim 5 or 6, wherein the SRD5A gene further comprises the SRD5A2 gene.

8. The method of any one of claims 1-4, wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.-59-4916-161 1 -5558.1Atty Dkt.: 064189-48909. The method of any one of claims 1, 2, or 3-8, wherein the non-hormonal therapy comprises one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibodydrug conjugates, or PARP inhibitors.

10. The method of any one of claims 1-9, wherein the chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

11. The method of any one of claims 1-7, 9 or 10, wherein the inherent variant is one or more at a position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100, optionally wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

12. The method of any one of claims 1-11, wherein the SRD5A genotype is determined by a method comprising one or more of PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods, optionally using a patient sample selected from a blood sample, a tissue sample such as a tumor sample.

13. The method of any one of claims 1-12, wherein the patient has undergone surgical resection of the prostate tumor.

14. The method of any one of claims 1-13, wherein the therapy is a first-line therapy.

15. A method for determine whether a prostate cancer patient who is naive to androgen ablation therapy is likely to be responsive to non-hormonal therapy, the method comprising determining whether a prostate tumor sample isolated from the patient contains a an inherent variant polymorphism in an SRD5A gene, wherein the patient is more likely to be responsive to non-hormonal therapy when the patient has the inherent variant polymorphism in the-60-4916-161 1 -5558.1Atty Dkt.: 064189-4890SRD5A gene and is less likely to be responsive to non-hormonal therapy when the patient lacks the inherent variant polymorphism or has wildtype genotype the SRD5A gene.

16. The method of claim 15, wherein the SRD5A gene is the SRD5A3 gene.

17. The method of claim 15 or 16, wherein the SRD5A gene comprises the SRD5A3 and the SRD5A1 genes.

18. The method of claim 15 or 16, wherein the SRD5A gene further comprises the SRD5A2 gene.

19. The method of any one of claims 15-18, wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

20. The method of any one of claims 15-19, wherein the non-hormonal therapy comprises one or more of chemotherapy, immunotherapy, radionucleotide therapy, antibody-drug conjugates, or PARP inhibitors.

21. The method of claim 20, wherein the chemotherapy comprises one or more of docetaxel, cabazitaxel, paclitaxel, mitoxantrone, cisplatin, carboplatin, oxaliplatin, gemcitabine, or Alimta.

22. The method of any one of claims 15-18, 21 or 22, wherein the inherent variant is one or more position selected from: SRD5A1 : rsl2716180, rsl042150, rs2434525, rs79421055, rs8192206, rsl6877779, rs562461, rs535981, rs8192249, rs28594474, rs4702381, rs329, rsl651071, rs3822430, rs248805, rs8192139, rs477930, 19 in SRD5A2: rs551741548, rs74403699, rs28383087, rs28383082, rs28383083, rs28383086, rs72794634, rs9332975, rsl92604242, rs781063043, rs6727380, rs28383085, rs2268796, rsl2470143, rs559555, rs2300697, rs632148, rsl988909, rs2366063, and 5 in SRD5A3: rs73236134, rs7674388, rs73236133, rs7663650, rs7664100, optionally wherein the inherent variant in the SRD5A gene is rs632148 in SRD5A2.

23. The method of any one of claims 15-22, wherein the SRD5A genotype is determined by a method comprising one or more of PCR, hybridization, whole genome sequencing, or TaqMan® SNP assay following manufacturers methods.-61-4916-161 1 -5558.1Atty Dkt.: 064189-489024. The method of any one of claims 15-23, wherein the patient has undergone surgical resection of the prostate tumor.

25. The method of any one of claims 15-24, wherein the therapy is first-line therapy.4916-161 1 -5558.1

Images