Bladder Cancer Treatment Methods

Abstract

Methods for treating bladder cancer are provided. [Solution] The present application provides methods, compositions (including pharmaceutical compositions), and kits for treating bladder cancer in an individual, comprising intravesical administration of an oncolytic virus, e.g., CG0070. In one aspect, provided herein is a method of treating bladder cancer in an individual, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus once a week for three weeks during a maintenance phase, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for oncolytic virus replication and a heterologous gene encoding an immune-related molecule.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 485,805, filed April 14, 2017, and U.S. Provisional Patent Application No. 62 / 500,729, filed May 3, 2017, all of which are incorporated by reference in their entireties.

[0002] The present invention relates to methods of treating bladder intraepithelial carcinoma using oncolytic viruses such as CG0070. [Background technology]

[0003] In 2016, approximately 77,000 new cases of bladder cancer were diagnosed. Non-muscle invasive bladder cancer (stage Ta, T1, or carcinoma in situ) accounts for 70-80% of these cases, while muscle invasive disease (stage T2 or higher) and metastatic disease make up the remaining 20-30%. Bladder cancer is classified into two subtypes, papillary and squamous, based on distinct patterns of cellular growth. Carcinoma in situ is a squamous tumor that is confined to the surface layer of the bladder. Compared to stage Ta and T1 papillary tumors, bladder CIS is more difficult to diagnose and carries a higher risk of progression to muscle invasive bladder cancer.

[0004] Intravesical Bacillus Calmette-Guerin (BCG) is the standard treatment for bladder CIS. Patients typically receive initial treatment consisting of weekly intravesical BCG instillations for 6 weeks, followed by monthly maintenance treatments for 6 to 12 months. However, approximately 30 to 40% of patients do not respond to a single BCG treatment. In one study, the overall risk of progression was approximately 14% in BCG-treated CIS patients with a median follow-up of 2.5 years. See Witjes, JA European urology 45.2 (2004): 142-146. Because the likelihood of progression in CIS patients who fail BCG is significant, cystectomy, including partial and radical cystectomy, remains a treatment option. However, cystectomy is associated with severe side effects and adversely affects the patient's quality of life. Furthermore, the risk of recurrence after radical cystectomy for clinically localized bladder cancer is high and stage-dependent. See, eg, Bassi P et al. J. Urol. 1999;161:1494-7. Elderly patients and those with renal failure tolerate surgery and chemotherapy less well, posing additional clinical challenges for the treatment of bladder CIS. The disclosures of all publications, patents, patent applications, and patent application publications referenced herein are hereby incorporated by reference in their entirety. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Witjes, JAEuropean urology 45.2(2004):142-146 [Non-Patent Document 2] Bassi P et al.J.Urol.1999;161:1494-7 Summary of the Invention [Means for solving the problem]

[0006] The present application provides methods, compositions (including pharmaceutical compositions), and kits for treating bladder cancer in an individual, comprising intravesical administration of an oncolytic virus, such as CG0070.

[0007] In one aspect, provided herein is a method of treating bladder cancer in an individual, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus once a week for three weeks during a maintenance phase, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule.

[0008] In another aspect, provided herein is a method of preserving the bladder in an individual, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus once a week for three weeks during a maintenance phase, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule.

[0009] In some embodiments, by any of the above methods, the oncolytic virus is administered once a week for three weeks every six months during the maintenance phase. In some embodiments, the methods of the invention further comprise an induction phase prior to the maintenance phase, the induction phase comprising administering an effective amount of the oncolytic virus to the individual once a week for six weeks. In some embodiments, the start of the induction phase and the start of the maintenance phase are separated by about three months. In some embodiments, the start of the induction phase and the start of the maintenance phase are separated by about six months. In some embodiments, the induction phase comprises administering an effective amount of the oncolytic virus to the individual once a week for six weeks at months 0 and 3 of the treatment regimen.

[0010] Also provided herein is a method for treating Ta or T1 bladder cancer in an individual who has not undergone transurethral resection of the bladder tumor (TURBT), the method comprising intravesically administering to the individual an effective amount of an oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule.

[0011] In one aspect of the present application, a method of treating bladder cancer in an individual (e.g., a human) is provided, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus, the oncolytic virus comprising a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the individual has only bladder carcinoma in situ. In some embodiments, the individual does not have concomitant papillary carcinoma at stage Ta or T1. In some embodiments, the individual has bladder carcinoma in situ and Ta or stage Ta cancer (CIS+Ta or CIS+T1). In some embodiments, the individual has Ta or T1 bladder cancer but not carcinoma in situ. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone a transurethral resection of the bladder tumor (TURBT). In some embodiments, the individual has unresectable stage Ta or T1 bladder cancer. In some embodiments, the individual does not respond to BCG treatment. In some embodiments, the individual has disease recurrence after BCG treatment. In some embodiments, the individual has failed BCG treatment within about 6 months. In some embodiments, the individual is BCG refractory.

[0012] In some embodiments according to any of the above methods, the individual has not undergone a cystectomy. In some embodiments, the individual has refused or is not suitable for a cystectomy. In some embodiments, the cystectomy is a radical cystectomy.

[0013] In some embodiments of any of the above methods, the oncolytic virus replicates preferentially in cancer cells. In some embodiments, the cancer cells have a defect in the Rb pathway. In some embodiments, the tumor-specific promoter is the E2F-1 promoter. In some embodiments, the E2F-1 promoter comprises the nucleotide sequence shown in SEQ ID NO:1.

[0014] In some embodiments according to any of the above methods, the immune related molecule is selected from the group consisting of GM-CSF, IL-2, IL-12, interferon, CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTαβ. In some embodiments, the immune related molecule is GM-CSF. In some embodiments, the heterologous gene is operably linked to a viral promoter.

[0015] In some embodiments of any of the above methods, the oncolytic virus is selected from the group consisting of adenovirus, herpes simplex virus, vaccinia virus, mumps virus, Newcastle disease virus, poliovirus, measles virus, Seneca Valley virus, coxsackievirus, reovirus, vesicular stomatitis virus, Maraba and rhabdovirus, and parvovirus. In some embodiments, the oncolytic virus is an oncolytic adenovirus. In some embodiments, the viral gene essential for the replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4. In some embodiments, the heterologous gene is operably linked to an E1 promoter or an E3 promoter. In some embodiments, the oncolytic virus is an adenovirus serotype 5, and the endogenous E1a promoter of the native adenovirus is replaced with a human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus is replaced with a nucleic acid encoding human GM-CSF. In some embodiments, the oncolytic virus is CG0070.

[0016] In some embodiments according to any of the above methods, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 It is administered in doses of viral particles.

[0017] In some embodiments of any of the above methods, the oncolytic virus is administered weekly, hi some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks.

[0018] In some embodiments according to any of the above methods, the method further comprises intravesically administering to the individual a transduction enhancing agent prior to administration of the oncolytic virus, hi some embodiments, the transduction enhancing agent is N-dodecyl-β-D-maltoside (DDM).

[0019] In some embodiments according to any of the above methods, the method is repeated at least one time.

[0020] In some embodiments according to any of the above methods, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent.

[0021] In some embodiments according to any of the above methods, the oncolytic virus is administered as the sole therapeutic agent.

[0022] Also, in one aspect, a pharmaceutical composition is provided for treating bladder cancer in an individual (e.g., a human), the pharmaceutical composition being administered intravesically to the individual, the pharmaceutical composition comprising an oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.

[0023] In a further aspect, there is provided a use of an oncolytic virus in the preparation of a medicament for treating bladder cancer in an individual (e.g., a human), wherein the medicament is administered intravesically to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.

[0024] These and other aspects and advantages of the present invention will become apparent from the following detailed description and the appended claims. It is to be understood that one, some, or all of the features of the various embodiments described herein may be combined to form other embodiments of the present invention. In an embodiment of the present invention, for example, the following items are provided: (Item 1) 2. A method for treating bladder intraepithelial carcinoma in an individual, comprising intravesically administering to the individual an effective amount of an oncolytic virus, the oncolytic virus comprising a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. 2. The method of claim 1, wherein the individual has bladder intraepithelial carcinoma only. (Item 3) 3. The method of claim 2, wherein the individual does not have stage Ta or T1 papillary carcinoma. (Item 4) 4. The method according to any one of items 1 to 3, wherein the oncolytic virus is administered once a week. (Item 5) 5. The method according to any one of items 1 to 4, wherein the oncolytic virus is administered for about 1 week to about 6 weeks. (Item 6) 6. The method according to any one of items 1 to 5, wherein the method is repeated at least one time. (Item 7) A method of treating bladder cancer in an individual, comprising administering to the individual an effective amount of an oncolytic virus intravesically once a week for three weeks during a maintenance phase, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. (Item 8) 1. A method of preserving the bladder in an individual, comprising administering to the individual an effective amount of an oncolytic virus intravesically once a week for three weeks during a maintenance phase, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. (Item 9) 9. The method of item 7 or 8, wherein the oncolytic virus is administered once a week for 3 weeks every 6 months during the maintenance phase. (Item 10) 10. The method of claim 9, further comprising an induction phase prior to said maintenance phase, said induction phase comprising administering to said individual an effective amount of an oncolytic virus once a week for six weeks. (Item 11) 11. The method of claim 10, wherein the start of the induction phase and the start of the maintenance phase are separated by about 3 months. (Item 12) 11. The method of claim 10, wherein the start of the induction phase and the start of the maintenance phase are separated by about 6 months. (Item 13) 11. The method of claim 10, wherein the induction phase comprises administering to the individual an effective amount of an oncolytic virus once a week for six weeks in months 0 and 3 of the treatment regimen. (Item 14) 14. The method according to any one of items 7 to 13, wherein the individual has carcinoma in situ. (Item 15) 15. The method of claim 14, wherein the individual does not have stage Ta or T1 papillary carcinoma. (Item 16) 15. The method of any one of items 7 to 14, wherein the individual has stage Ta or T1 bladder cancer. (Item 17) A method for treating Ta or T1 bladder cancer in an individual who has not undergone transurethral resection of the bladder tumor (TURBT), comprising intravesically administering to the individual an effective amount of an oncolytic virus, the oncolytic virus comprising a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. (Item 18) 18. The method according to any one of items 1 to 17, wherein the individual is non-responsive to BCG treatment. (Item 19) 18. The method according to any one of items 1 to 17, wherein the individual has disease recurrence after BCG treatment. (Item 20) 20. The method of item 18 or 19, wherein the individual has failed BCG treatment within about 6 months. (Item 21) 21. The method of any one of items 1 to 20, wherein the individual has not undergone a cystectomy. (Item 22) 22. The method of claim 21, wherein the individual has refused or is not suitable for cystectomy. (Item 23) 23. The method according to any one of items 1 to 22, wherein the oncolytic virus replicates preferentially in cancer cells. (Item 24) 24. The method of claim 23, wherein the cancer cells have a defect in the Rb pathway. (Item 25) 24. The method of claim 23, wherein the tumor-specific promoter is an E2F-1 promoter. (Item 26) 26. The method of claim 25, wherein the E2F-1 promoter comprises the nucleotide sequence set forth in SEQ ID NO:1. (Item 27) 27. The method according to any one of items 1 to 26, wherein the immune-related molecule is selected from the group consisting of GM-CSF, IL-2, IL-12, interferon, CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTαβ. (Item 28) 28. The method of claim 27, wherein the immune-related molecule is GM-CSF. (Item 29) 29. The method according to any one of items 1 to 28, wherein the heterologous gene is operably linked to a viral promoter. (Item 30) 30. The method according to any one of items 1 to 29, wherein the oncolytic virus is selected from the group consisting of adenovirus, herpes simplex virus, vaccinia virus, mumps virus, Newcastle disease virus, poliovirus, measles virus, Seneca Valley virus, coxsackievirus, reovirus, vesicular stomatitis virus, Maraba and rhabdovirus, and parvovirus. (Item 31) 31. The method of item 30, wherein the oncolytic virus is an oncolytic adenovirus. (Item 32) 32. The method of claim 31, wherein the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4. (Item 33) 31. The method of claim 29 or 30, wherein the heterologous gene is operably linked to an E1 promoter or an E3 promoter. (Item 34) 34. The method according to any one of items 1 to 33, wherein the oncolytic virus is adenovirus serotype 5, the endogenous E1a promoter of the native adenovirus is replaced with a human E2F-1 promoter, and the endogenous E3 19 kD coding region of the native adenovirus is replaced with a nucleic acid encoding human GM-CSF. (Item 35) 35. The method of item 34, wherein the oncolytic virus is CG0070. (Item 36) The oncolytic virus is about 1×10 8 ~Approx. 1×10 14 36. The method of any one of items 1 to 35, wherein the viral particles are administered in a dose of 100 μg / kg. (Item 37) 37. The method of any one of items 1 to 36, wherein the oncolytic virus is administered in a volume of 50 mL. (Item 38) 38. The method of any one of items 1 to 37, further comprising intravesically administering a transduction enhancing agent to the individual prior to administration of the oncolytic virus. (Item 39) 39. The method of claim 38, wherein the transduction enhancing agent is N-dodecyl-β-D-maltoside (DDM). (Item 40) 40. The method of any one of items 1 to 39, wherein the oncolytic virus is not administered in combination with an immune checkpoint modulator. (Item 41) 41. The method of any one of items 1 to 40, wherein the oncolytic virus is administered as a single therapeutic agent. (Item 42) 42. The method according to any one of items 1 to 41, wherein the individual is a human. [Brief description of the drawings]

[0025] [Figure 1] Schematic diagram of CG0070 and wild-type (wt) adenovirus type 5. CG0070 is based on adenovirus serotype 5, but the endogenous E1a promoter and E3 19 kD coding region have been replaced with the human E2F-1 promoter and cDNA coding region for human GM-CSF, respectively.

[0026] [Diagram 2] Figure 1 shows complete response (CR) rates in patients with various stages of high-grade non-muscle invasive bladder cancer 6 months after receiving intravesical CG0070 treatment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] The present invention provides methods and compositions for treating bladder cancer in an individual by intravesically administering an effective amount of an oncolytic virus, such as CG0070, to the individual. In vitro studies suggest that the use of conditionally replicating oncolytic viruses, such as CG0070, may be an effective strategy for the treatment of bladder cancer. Ramesh N, Ge Y, Ennist DL, Zhu M, Mina M, Ganesh S, Reddy PS, Yu DC. CG0070, a conditionally replicating granulocyte macrophage colony-stimulating factor-armed oncolytic adenovirus for the treatment of bladder cancer. Clin Cancer Res. 2006; 12: 305-13. However, the unexpected efficacy of CG0070 in certain patient populations, and the effect of the specific dosing regimen described herein, was previously unknown. The method is bladder-preserving, highly effective, and well tolerated in bladder cancer, including those that have previously failed standard therapies. In a Phase II clinical trial, the inventors surprisingly found that 72.2% of patients with pure CIS who had previously failed BCG therapy achieved a complete response (CR) after 6 months of intravesical CG0070 treatment. The therapeutic effect of oncolytic viruses can be achieved using oncolytic viruses as monotherapy, i.e., without combining with immune checkpoint modulators. The methods and compositions described herein are particularly useful for patients with high-risk bladder CIS who refuse or are not suitable for cystectomy.

[0028] Thus, one aspect of the present application provides a method of treating bladder cancer in an individual, the method comprising intravesically administering an effective amount of an oncolytic virus to the individual. In some embodiments, the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus is CG0070. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulator. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment.

[0029] Also provided are compositions (such as pharmaceutical compositions), medicaments, kits, and unit dosages that are useful for the methods described herein.

[0030] I. Definition Terms are used herein as commonly used in the art unless otherwise defined below.

[0031] As used herein, "bladder intraepithelial carcinoma," "bladder CIS," and "carcinoma in situ of the bladder" are used interchangeably and refer to clinical stages of bladder cancer characterized by squamous (i.e., non-papillary) lesions composed of cytologically malignant cells that may involve either full or partial thickness of the urothelium.

[0032] As used herein, "treatment" or "treating" is a method for obtaining beneficial or desired results, including clinical results. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms resulting from bladder cancer, reducing the extent of bladder cancer, stabilizing bladder cancer (e.g., preventing or slowing the progression of bladder cancer), stabilizing bladder cancer (e.g., preventing or slowing the spread of bladder cancer (e.g., metastasis), preventing or slowing the recurrence of bladder cancer, reducing the recurrence rate of bladder cancer, slowing or slowing the progression of bladder cancer, remission of bladder cancer condition, causing remission (partial or complete) of bladder cancer, reducing the dose of one or more other medications required to treat bladder cancer, slowing the progression of bladder cancer, improving quality of life, and / or increasing survival. "Treatment" also encompasses the reduction of pathological consequences of bladder cancer. The methods of the present invention contemplate any one or more of these aspects of treatment.

[0033] As used herein, "prior treatment" refers to a treatment regimen that is distinct from and that is initiated prior to the methods described herein, including intravesical administration of an oncolytic virus.

[0034] As used herein, an "at risk" individual is a human individual at risk of developing bladder CIS. An "at risk" human individual may or may not have detectable disease, or may or may not exhibit detectable disease prior to the treatment methods described herein. "At risk" indicates that a human individual has one or more so-called risk factors, which are measurable parameters that correlate with the development of muscle-invasive bladder cancer, as described herein. A human individual who has one or more of these risk factors has a higher probability of developing bladder CIS than a human individual who does not have these risk factors.

[0035] "Adjuvant setting" refers to a clinical setting in which an individual has a history of bladder cancer and has usually (but not necessarily) benefited from treatment, including, but not limited to, surgical treatment (e.g., transurethral resection of the bladder tumor (TURBT), partial cystectomy, radical cystectomy), radiation therapy, and chemotherapy. Treatment and administration in the "adjuvant setting" refers to the mechanism of treatment that follows.

[0036] "Neoadjuvant setting" refers to the clinical setting in which the method is administered prior to primary / definitive treatment.

[0037] The terms "individual," "subject," and "patient" are used interchangeably herein to refer to a mammal, including a human. An individual includes, but is not limited to, a human, a bovine, an equine, a feline, a canine, a rodent, or a primate. In some embodiments, the individual is a human. In some embodiments, the individual suffers from bladder CIS. In some embodiments, the individual is in need of treatment.

[0038] As used herein, "delaying" the onset of bladder cancer means delaying, preventing, slowing, inhibiting, stabilizing, and / or postponing the onset of bladder cancer. This delay can be of various durations, depending on the history of the disease and / or the individual being treated. As will be apparent to those skilled in the art, a sufficient or significant delay may actually encompass prevention, in that the individual does not develop the disease. A method of "delaying" the onset of bladder cancer is one that reduces the probability of the disease developing for a period of time and / or reduces the extent of bladder cancer for a period of time, compared to when the method is not used. Such comparisons are typically based on clinical trials using a statistically significant number of subjects. The onset of bladder cancer can be detected using standard methods, including, but not limited to, urine cytology, urethral cystoscopy (UCS), computed tomography (CT scan, e.g., spiral CT scan), endoscopic ultrasound (EUS), endoscopic retrograde cholangiopancreatography (ERCP), laparoscopy, or biopsy (e.g., percutaneous needle biopsy or fine needle aspiration). Onset can also refer to the progression of bladder cancer that is not detectable in the early stages, including recurrence.

[0039] As used herein, "combination therapy" means that a first agent is administered in combination with another agent. "Combination" refers to administering one therapeutic modality in addition to another therapeutic modality. That is, "combination" refers to administering one therapeutic modality before, during, or after the delivery of another therapeutic modality to an individual.

[0040] As used herein, "monotherapy" refers to the administration of a single therapeutic agent, such as an oncolytic virus, and not in combination with another therapeutic modality, such as an immune checkpoint modulator. Oncolytic virus monotherapy can be administered with a pretreatment agent, such as a transduction enhancing agent (e.g., DDM), which is not considered a distinct therapeutic modality for purposes of the present invention.

[0041] As used herein, the term "effective amount" refers to an amount of a compound (e.g., an oncolytic virus described herein) or composition sufficient to treat a specified disorder, condition, or disease, such as ameliorating, alleviating, reducing, and / or delaying one or more of the symptoms. In the context of cancer, an effective amount includes an amount sufficient to shrink a tumor and / or reduce the rate of tumor growth (e.g., inhibit tumor growth) or prevent or slow other unwanted cell proliferation in cancer. In some embodiments, an effective amount is an amount sufficient to slow the progression of bladder cancer. In some embodiments, an effective amount is an amount sufficient to prevent or slow recurrence. In some embodiments, an effective amount is an amount sufficient to reduce the recurrence rate in an individual. In some embodiments, an effective amount is an amount sufficient to inhibit tumor metastasis in an individual. An effective amount may be administered in one or more administrations. An effective amount of an agent or composition may (i) reduce the number of cancer cells, (ii) reduce tumor size, (iii) inhibit, delay, slow to some extent, and preferably stop, cancer cell invasion into peripheral organs, (iv) inhibit (i.e., slow to some extent, and preferably stop) tumor metastasis, (v) inhibit tumor growth, (vi) prevent tumor onset and / or recurrence, (vii) delay tumor onset and / or recurrence, (viii) reduce the recurrence rate of tumors, and / or (ix) relieve to some extent one or more of the symptoms associated with cancer. As is understood in the art, an "effective amount" may be in one or more administrations, i.e., a single administration or multiple administrations may be required to achieve the desired therapeutic endpoint.

[0042] As used herein, the term "co-administration" means that a first and second treatment as a combination therapy are administered at the same time. When a first and second treatment are administered at the same time, the first and second treatments may be contained in the same composition (e.g., a composition that includes both the first and second treatments) or may be contained in separate compositions (e.g., the first treatment is contained in one composition and the second treatment is contained in another composition).

[0043] As used herein, the term "sequential administration" or "in order" means that the first and second treatments of the combination therapy are administered more than about one minute apart, such as more than about 5, 10, 15, 20, 30, 40, 50, 60 minutes, or more. In some cases, the term "sequential administration" means that the first and second treatments of the combination therapy are administered more than about one day apart, such as more than about one day to one week, two weeks, three weeks, four weeks, eight weeks, twelve weeks, or more weeks. Either the first or second treatment may be administered first. The first and second treatments are contained in separate compositions, which may be included in the same or separate packages or kits.

[0044] The term "administered immediately before" means that the first treatment is administered within about 15 minutes, such as within about any of 10, 5, or 1 minutes, before the administration of the second treatment. The term "administered immediately after" means that the first treatment is administered within about 15 minutes, such as within about any of 15, 10, or 1 minutes, after the administration of the second treatment.

[0045] As used herein, "pharmacologically acceptable" or "pharmacologically compatible" means a substance that is not biologically or non-biologically undesirable, e.g., the substance can be incorporated into a pharmaceutical composition administered to a patient without causing significant undesired biological effects or adversely interacting with any other components of the composition in which it is contained. A pharma-ceutically acceptable carrier or excipient preferably meets the necessary toxicological and manufacturing testing standards and / or is included in the Inactive Ingredient Guide prepared by the US Food and Drug administration.

[0046] As used herein, "adverse event" or "AE" refers to any untoward medical occurrence in an individual receiving a marketed medicinal product or in an individual participating in a clinical trial receiving an investigational or non-investigational product. An AE does not necessarily have a causal relationship to the treatment of an individual. Thus, an AE can be any untoward, unintended sign, symptom, or disease that is temporally related to the use of a medicinal product, whether or not it is causally related to the medicinal product. AEs include, but are not limited to, exacerbation of a pre-existing disease, an increase in the frequency or intensity of a pre-existing episodic event or condition, a condition detected or diagnosed after administration of an investigational product (even if present before the start of the study), and a continuing, persistent disease or symptom that was present at the start of the study and worsened after the start of the study. Typically, AEs do not include medical or surgical procedures (e.g., surgery, endoscopy, tooth extraction, or blood transfusion) (although the condition leading to the procedure is an adverse event); pre-existing diseases, conditions, or laboratory abnormalities that were present or detected at the start of the study and have not worsened; hospitalizations or procedures performed for elective purposes not related to the adverse medical event (e.g., hospitalization for cosmetic or elective surgery, or social / convenience hospitalization); signs / symptoms of the disease being studied or related to that disease, unless the individual's condition is more severe than expected; and overdose of investigational drug without any clinical signs or symptoms.

[0047] As used herein, "serious adverse event" or (SAE) refers to any untoward medical occurrence at any dose, including, but not limited to, a) resulting in death; b) being life threatening (defined as imminent risk of death from the event that occurred); c) resulting in long-term or significant disability or incapacity; d) requiring hospitalization or prolonged hospitalization (Exception: Hospitalization for elective treatment of a pre-existing condition that did not worsen during the study is not considered an adverse event. Complications occurring during hospitalization are AEs, and if the complication prolongs the hospitalization, the event is serious); e) congenital malformation / birth defect in the offspring of the individual administered the drug; or f) a condition not included in the above definition that may endanger the individual or require intervention to prevent one of the above outcomes, except when clearly related to the individual's primary disease. "Lack of efficacy" (progression) is not considered an AE or SAE. Signs and symptoms attributable to lack of efficacy or clinical sequelae must be reported if they meet the definition of an AE or SAE.

[0048] The following definitions can be used to assess response based on target lesions: "complete response" or "CR" refers to disappearance of all target lesions; "partial response" or "PR" refers to a reduction in the sum of the longest diameters (SLD) of the target lesions of at least 30% compared to baseline SLD; "stable disease" or "SD" refers to insufficient reduction in target lesions for PR and insufficient growth for PD compared to the smallest SLD since treatment initiation; and "progression" or "PD" refers to an increase in the SLD of target lesions of at least 20% compared to the smallest SLD recorded since treatment initiation or the presence of one or more new lesions.

[0049] Non-target lesions can be assessed using the following response definitions: "complete response" or "CR" refers to disappearance of all non-target lesions; "stable disease" or "SD" refers to persistence of one or more non-target lesions that are not CR or PD; "progression" or "PD" refers to "clear worsening" of existing non-target lesions, with the presence of one or more new lesions being considered progression (if PD in an individual is assessed for a time point solely on the basis of progression of non-target lesions, additional criteria are required to perform the assessment).

[0050] "Progression-free survival" (PFS) refers to the time during and after treatment during which cancer does not grow. Progression-free survival includes time during which an individual has a complete or partial response, as well as time during which an individual has stable disease.

[0051] "Cystectomy-free survival" (CFS) refers to the period during and after treatment during which a patient does not require a cystectomy, as determined by a physician.

[0052] "Predict" or "prediction" is used herein to refer to the likelihood of responding, either favorably or unfavorably, to a therapeutic regimen.

[0053] As used herein, "initiation of treatment" or "baseline" refers to the time at or prior to first exposure to treatment.

[0054] As used herein, a "sample" refers to a composition containing molecules that are to be characterized and / or identified, e.g., based on physical, biochemical, chemical, physiological, and / or genetic characteristics.

[0055] It will be understood that embodiments of the invention described herein include "consisting of" and / or "consisting essentially of" embodiments.

[0056] Reference to "about" a value or parameter described herein includes (and describes) variations that are directed to the value or parameter itself. For example, a reference to "about X" includes a description of "X."

[0057] As used herein, reference to a value or parameter with a negation (not), such as "is not," generally means and describes the affirmative "other" of that value or parameter. For example, "the method is not used to treat cancer of type X" means "the method is used to treat cancers of types other than X."

[0058] As used herein, the term "about X to Y" has the same meaning as "about X to about Y."

[0059] As used in this specification and the appended claims, the singular forms "a," "or," and "the" are intended to include the plural of the referent unless the context clearly dictates a singular referent.

[0060] II. Treatment methods for bladder cancer One aspect of the present application relates to the treatment of bladder cancer in an individual (e.g., a human individual) by local administration of a virus, such as an oncolytic virus. In this context, local administration of an oncolytic virus includes intravesical administration of an oncolytic virus. Any of the methods described herein may be useful for inhibiting bladder tumor growth, inhibiting bladder tumor metastasis, prolonging the survival (e.g., disease-free survival, progression-free survival, or cystectomy-free survival) of an individual with bladder cancer, inducing disease remission in an individual with bladder cancer, preventing disease progression in an individual with bladder cancer, and / or improving the quality of life of an individual with bladder cancer. In some embodiments, the method is bladder preservation or bladder sparing. In some embodiments, bladder preservation or bladder sparing methods are useful for improving the quality of life of an individual.

[0061] In some embodiments, provided herein are methods of treating bladder cancer, comprising intravesically administering an oncolytic virus (e.g., CG0070) to an individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule, wherein the oncolytic virus is administered once a week for three weeks. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is delivered once a week for six weeks at month 0 and once a week for three weeks at month 6.

[0062] In some embodiments, the oncolytic virus is administered during the induction and maintenance phases. In some embodiments, the induction period comprises administering the oncolytic virus once a week for six weeks. In some embodiments, the induction period comprises administering the oncolytic virus once a week for six weeks at months 0 and 3.

[0063] In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for three weeks following the induction phase. In some embodiments, the maintenance phase comprises delivering the oncolytic virus once a week for three weeks every six months. In some embodiments, the maintenance phase comprises administering the oncolytic virus 2, 3, 4, 5, 6 times, or as needed, once a week for three weeks every six months. In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for three weeks at months 6, 12, and 18. In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for three weeks at months 3, 6, 12, and 18.

[0064] In some embodiments, the administration schedule can be adjusted based on the individual's response to the oncolytic virus.For example, in some embodiments, the individual is administered the oncolytic virus once a week for 6 weeks at month 0 and reassessed at month 3.In some embodiments, the individual who achieves a complete response at month 3 begins a maintenance phase that includes administering the oncolytic virus once a week for 6 weeks every 6 months.In some embodiments, the individual who does not achieve a complete response at month 3 receives a second induction administration of the oncolytic virus once a week for 6 weeks.

[0065] Thus, provided herein is a method of treating bladder cancer in an individual, the method comprising administering an oncolytic virus once a week for three weeks every six months, wherein the oncolytic virus (e.g., CG0070) is administered intravesically, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, CIS without Ta or T1, Ta or T1 grade bladder cancer. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone transurethral resection of the bladder tumor (TURBT). In some embodiments, the bladder cancer is Ta or T1 unresectable bladder cancer. In some embodiments, the oncolytic virus is administered at months 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered at months 3, 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered once a week for three weeks every six months as needed.

[0066] Thus, provided herein is a method of treating bladder cancer, comprising administering an oncolytic virus once a week for six weeks during an induction phase and once a week for three weeks during a maintenance phase, wherein the oncolytic virus (e.g., CG0070) is administered intravesically, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the bladder cancer is a bladder cancer of grade CIS, CIS+Ta, CIS+T1, CIS without Ta or T1, Ta or T1. In some embodiments, the bladder cancer is a Ta or T1 non-respectable bladder cancer. In some embodiments, the induction phase comprises administering the oncolytic virus once a week for 6 weeks at months 0 and 3. In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for 3 weeks at months 6, 12, and 18. In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for 3 weeks at months 3, 6, 12, and 18. In some embodiments, the maintenance phase comprises administering the oncolytic virus once a week for 3 weeks every 6 months as needed.

[0067] The method has the advantage of reducing the need for cystectomy in patients, and thus can be used in bladder-preserving treatment to treat patients who are not suitable for cystectomy. Thus, in some embodiments, a method of bladder-preserving treatment in an individual is provided herein, comprising administering an oncolytic virus once a week for three weeks every six months, wherein the oncolytic virus (e.g., CG0070) is administered intravesically, where the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for the replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor-selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulator. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, CIS without Ta or T1, Ta or T1 grade bladder cancer. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone transurethral resection of the bladder tumor (TURBT). In some embodiments, the bladder cancer is Ta or T1 unresectable bladder cancer. In some embodiments, the oncolytic virus is administered at months 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered at months 3, 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered once a week for three weeks every six months as needed.

[0068] In some embodiments, provided herein is a method of maintenance therapy, comprising administering an oncolytic virus once a week for three weeks every six months, wherein the oncolytic virus (e.g., CG0070) is administered intravesically, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, CIS without Ta or T1, Ta or T1 grade bladder cancer. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone transurethral resection of the bladder tumor (TURBT). In some embodiments, the bladder cancer is Ta or T1 unresectable bladder cancer. In some embodiments, the oncolytic virus is administered at months 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered at months 3, 6, 12, and 18 of the treatment regimen. In some embodiments, the oncolytic virus is administered once a week for three weeks every six months as needed.

[0069] In some embodiments, the individual is administered the oncolytic virus no more than 21 times. In some embodiments, the individual is administered the oncolytic virus no more than 30, 25, 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, or 6 times.

[0070] In some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus (e.g., an oncolytic adenovirus) comprising a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the tumor selective promoter allows preferential replication of the oncolytic virus in tumor cells. In some embodiments, the oncolytic virus replicates preferentially in cancer cells, such as cancer cells with a deletion in the Rb pathway. In some embodiments, the oncolytic virus is selected from the group consisting of adenovirus, herpes simplex virus, vaccinia virus, mumps virus, Newcastle disease virus, poliovirus, measles virus, Seneca Valley virus, coxsackievirus, reovirus, vesicular stomatitis virus, maraba and rhabdovirus, and parvovirus. In some embodiments, the tumor-specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1. In some embodiments, the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4.

[0071] In some embodiments, the oncolytic virus further comprises an immune related molecule (such as a cytokine, chemokine, or PRRago (i.e., a pathogen recognition receptor agonist)). In some embodiments, the immune related molecule is not an immune checkpoint modulator. In some embodiments, the immune related molecule is selected from the group consisting of GM-CSF, IL-2, IL-12, interferon (such as type 1, type 2, or type 3 interferon, e.g., interferon gamma gamma), CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTααββ. In some embodiments, the immune-related molecule is selected from the group consisting of STING (i.e., stimulator of interferon genes) activators (such as CDN, i.e., cyclic dinucleotides), PRRago (such as CpG, imiquimod, or poly I:C), TLR stimulators (such as GS-9620, AED-1419, CYT-003-QbG10, AVE-0675, or PF-7909), and RLR stimulators (such as RIG-I, Mda5, or LGP2 stimulators). In some embodiments, the immune-related molecule attracts dendritic cells, T cells, B cells, and / or follicular helper T cells.

[0072] In some embodiments, the immune-related molecule is expressed by an oncolytic virus. For example, the oncolytic virus may comprise a nucleic acid encoding the immune-related molecule, which may be present in a viral vector or on a separate vector. In some embodiments, the oncolytic virus is a virus that comprises a viral vector, where the viral vector comprises a nucleic acid encoding the immune-related molecule. In some embodiments, the nucleic acid encoding the immune-related molecule is operably linked to a viral promoter, such as an E1 promoter or an E3 promoter.

[0073] In some embodiments, the immune-related molecules enhance the immune response in an individual. Immune-related molecules include cytokines, chemokines, stem cell growth factors, lymphotoxins, hematopoietic factors, colony-stimulating factors (CSFs), erythropoietin, thrombopoietin, tumor necrosis factor alpha (TNF), TNF-beta, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha, interferon beta, interferon gamma, interferon lambda, stem cell growth factor known as "S1 factor", human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), hepatic growth factor, prostaglandins, fibroblast growth factors, prolactin, and the like. In some embodiments, the therapeutic agent may include, but is not limited to, IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, FLT-3, angiostatin, thrombospondin, endostatin, lymphotoxin, thalidomide, lenalidomide, or pomalidomide.

[0074] Thus, in some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for oncolytic virus replication and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine) operably linked to the viral promoter. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the tumor specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the viral gene essential for oncolytic virus replication is selected from the group consisting of E1A, E1B, and E4. In some embodiments, the viral promoter operably linked to a nucleic acid encoding an immune-related molecule is an E3 promoter. In some embodiments, the immune-related molecule is GM-CSF. In some embodiments, the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 6 weeks, or 3 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0075] In some embodiments, the oncolytic virus is adenovirus serotype 5. In some embodiments, the endogenous E1a promoter and E3 19 kD coding region of the native adenovirus are replaced with the human E2F-1 promoter and a nucleic acid encoding human GM-CSF. In some embodiments, a polyadenylation signal (PA) is inserted 5' of the E2F-1 promoter. In some embodiments, the nucleic acid encoding human GM-CSF is operably linked to the E3 promoter. In some embodiments, the adenovirus serotype 5 vector backbone further comprises E2, E4, a late protein region, or an inverted terminal repeat (ITR) identical to the wild-type adenovirus serotype 5 genome. In some embodiments, the oncolytic virus has a genome structure as shown in FIG. 1. In some embodiments, the oncolytic virus is conditionally replicating. In some embodiments, the oncolytic virus grows preferentially in cancer cells. In some embodiments, the cancer cells are Rb pathway-deficient cancer cells. In some embodiments, the oncolytic virus is CG0070.

[0076] Thus, in some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, the method comprising intravesically administering to the individual an effective amount of adenovirus serotype 5, wherein the endogenous E1a promoter and E3 19 kD coding region of the native adenovirus are replaced with a human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (such as a cytokine or chemokine, e.g., GM-CSF). In some embodiments, the tumor-specific promoter is a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to administration of the adenovirus. In some embodiments, the adenovirus is administered to the individual at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the adenovirus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the adenovirus is administered once a week. In some embodiments, the adenovirus is administered for about 1 week to about 6 weeks (e.g., about 6 weeks, or 3 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0077] In some embodiments, methods of treating bladder cancer (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) are provided, the methods comprising intravesically administering an effective amount of CG0070 to an individual. In some embodiments, the methods further comprise administering to the individual a transduction enhancing agent (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, CG0070 is administered at a dose of 1000 to 2000 viral particles (viral particles). In some embodiments, CG0070 is administered once per week. In some embodiments, CG0070 is administered for about 1 week to about 6 weeks (e.g., about 6 weeks, or 3 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0078] In some embodiments, the oncolytic virus is not administered to an individual in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered to an individual as a single therapeutic agent. As used herein, an "immune checkpoint modulating agent" refers to a molecule or agent (e.g., an antibody) that inhibits an inhibitory immune checkpoint molecule or an immune stimulatory agent that activates an immune stimulatory molecule in an immune cell (e.g., T cell) or tumor cell. An "immune checkpoint molecule" includes a molecule that increases an immune signal (i.e., an "immunostimulatory molecule") or decreases an immune signal (i.e., an "inhibitory immune checkpoint molecule") to a tumor cell.

[0079] In some embodiments, the oncolytic virus is not administered to an individual in combination with an immunostimulatory agent. The immunostimulatory agent may be a natural or artificial ligand of an immunostimulatory molecule selected from the group consisting of OX40 ligand (e.g., OX40L), CD-28 ligand (e.g., CD80, CD86), ICOS ligand (e.g., B7RP1), 4-1BB ligand (e.g., 4-1BBL, Ultra4-1BBL), CD27 ligand (e.g., CD70), CD40 ligand (e.g., CD40L), and TCR ligand (e.g., MHC class I or class II molecule, IMCgp100). The immune stimulant may be an antibody selected from the group consisting of anti-CD28 (e.g., TGN-1412), anti-OX40 (e.g., MEDI6469, MEDI-0562), anti-ICOS (e.g., MEDI-570), anti-GITR (e.g., TRX518, INBRX-110, NOV-120301), anti-41-BB (e.g., BMS-663513, PF-05082566), anti-CD27 (e.g., BION-1402, varlilumab and hCD27.15), anti-CD40 (e.g., CP870, 893, BI-655064, BMS-986090, APX005, APX005M), anti-CD3 (e.g., blinatumomab, muromonab), and anti-HVEM.

[0080] In some embodiments, the oncolytic virus is not administered to the individual in combination with an immune checkpoint inhibitor. The immune checkpoint inhibitor may be a natural or artificial ligand of an inhibitory immune checkpoint molecule selected from the group consisting of a ligand of CTLA-4 (e.g., B7.1, B7.2), a ligand of TIM3 (e.g., Galectin-9), a ligand of A2a receptor (e.g., adenosine, regadenoson), a ligand of LAG3 (e.g., MHC class I or MHC class II molecule), a ligand of BTLA (e.g., HVEM, B7-H4), a ligand of KIR (e.g., MHC class I or MHC class II molecule), a ligand of PD-1 (e.g., PD-L1, PD-L2), a ligand of IDO (e.g., NKTR-218, Indoximod, NLG919), and a ligand of CD47 (e.g., SIRP-α receptor).Immune checkpoint inhibitors include anti-CTLA-4 (e.g., ipilimumab, tremelimumab, KAHR-102), anti-TIM3 (e.g., F38-2E2, ENUM005), anti-LAG3 (e.g., BMS-986016, IMP701, IMP321, C9B7W), anti-KIR (e.g., lirilumab and IPH2101), anti-PD-1 (e.g., nivolumab, pidilizumab, pembrolizumab, BMS-936559, atezol izumab, lambrolizumab, MK-3475, AMP-224, AMP-514, STI-A1110, TSR-042), anti-PD-L1 (e.g., KY-1003 (EP20120194977), MCLA-145, RG7446, BMS-936559, MEDI-4736, MSB0010718C, AUR-012, STI-A1010, PCT / US2001 / 020964, MPDL3280A, AMP-224, Dapirolizumab pegol (CDP-7657), MEDI-4920), anti-CD73 (e.g., AR-42 (OSU-HDAC42, HDAC-42, AR 42, AR42, OSU-HDAC 42, OSU-HDAC-42, NSC D736012, HDAC-42, HDAC 42, HDAC42, NSCD736012, NSC-D736012), MEDI-9447), anti-B7-H3 (e.g., MGA271, DS-5573a, 8H9), anti-CD47 (e.g., CC-90002, TTI-621, VLST-007), anti-BTLA, anti-VISTA, anti-A2aR, anti-B7-1, anti-B7-H4, anti-CD52 (e.g., Alemtuzumab), anti-IL-10, anti-IL-35, and anti-TGF-β (e.g., Fresolumimab).

[0081] Thus, in some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, comprising administering to the individual an effective amount of an oncolytic virus (e.g., an oncolytic adenovirus) comprising a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine) operably linked to the viral promoter, wherein the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, a method of treating bladder cancer in an individual is provided, the method comprising intravesically administering to the individual an effective amount of an oncolytic virus (e.g., an oncolytic adenovirus) comprising a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine) operably linked to the viral promoter, wherein the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the tumor specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4. In some embodiments, the viral promoter operably linked to a nucleic acid encoding an immune-related molecule is an E3 promoter. In some embodiments, the immune-related molecule is GM-CSF. In some embodiments, the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 6 weeks, or 3 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0082] In some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, the method comprising intravesically administering to the individual an effective amount of adenovirus serotype 5, wherein the endogenous E1a promoter and E3 19 kD coding region of the native adenovirus have been replaced with a human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (such as a cytokine or chemokine, e.g., GM-CSF), and the adenovirus is administered as the single therapeutic agent. In some embodiments, a method of treating bladder intraepithelial carcinoma in an individual is provided, the method comprising intravesically administering to the individual an effective amount of adenovirus serotype 5, wherein the endogenous E1a promoter and E3 19 kD coding region of the native adenovirus are replaced with a human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (such as a cytokine or chemokine, e.g., GM-CSF), and the adenovirus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the tumor-specific promoter is a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to administration of the adenovirus. In some embodiments, the adenovirus is administered in a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the adenovirus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the adenovirus is administered once a week. In some embodiments, the adenovirus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0083] In some embodiments, a method of treating bladder cancer in an individual (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, the method comprising intravesically administering an effective amount of CG0070 to the individual, where CG0070 is administered as a single therapeutic agent. In some embodiments, a method of treating bladder intraepithelial carcinoma in an individual is provided, the method comprising intravesically administering an effective amount of CG0070 to the individual, where CG0070 is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of CG0070. In some embodiments, CG0070 is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, CG0070 is administered at a dose of 1000 to 2000 viral particles (viral particles). In some embodiments, CG0070 is administered once per week. In some embodiments, CG0070 is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0084] Patient population The methods described herein can be used to treat various bladder cancers. In some embodiments, the individual has bladder cancer at stage Ta, T1, or CIS (TIS) as determined by the TNM staging system according to the American Joint Committee on Cancer (AJCC) guidelines.

[0085] In some embodiments, the bladder cancer is CIS. In some embodiments, the bladder CIS is transitional cell carcinoma or urothelial carcinoma. In some embodiments, the bladder CIS is metastatic urothelial carcinoma. In some embodiments, the bladder CIS is located in the urothelium of the bladder. In some embodiments, the bladder CIS is located in the upper urinary tract. In some embodiments, the bladder CIS is in the ureter. In some embodiments, the bladder CIS is in the urethra. In some embodiments, the bladder CIS is in the renal pelvis.

[0086] "Bladder intraepithelial carcinoma", also known as stage 0is, Tis, squamous, or bladder CIS, is an aggressive form of bladder cancer. Bladder CIS is found only in the inner wall of the bladder, has not grown toward the bladder cavity, and has not spread into the thick muscular layer of the bladder or the connective tissue of the bladder. Bladder CIS is considered an aggressive disease and is thought to be a precursor to invasive bladder cancer. According to the NCCN guidelines for bladder cancer, the current standard of care for bladder CIS is resection, followed by intravesical treatment with BCG. BCG is generally administered weekly for 6 weeks, followed by a 4-6 week break, and is fully reevaluated at 12 weeks after starting treatment. If the patient cannot tolerate BCG, mitomycin C may be administered intravesically.

[0087] In some embodiments, the individual has a pathological stage of "pure CIS" or "CIS", where the individual has one or more sites of bladder intraepithelial carcinoma. In some embodiments, the individual has a pathological stage of Tis, N0, M0. In some embodiments, the bladder CIS is a primary tumor. In some embodiments, the bladder CIS is a recurrent tumor. In some embodiments, the individual has at least about any of the following: 1, 2, 3, 4, 5, 10, or more sites of bladder intraepithelial carcinoma.

[0088] In some embodiments, the individual has, simultaneously, one or more sites of bladder intraepithelial carcinoma and one or more sites of stage Ta or T1 papillary carcinoma. In some embodiments, the individual has a pathological stage of "CIS+Ta", where the individual has, simultaneously, one or more sites of bladder intraepithelial carcinoma and one or more sites of stage Ta papillary carcinoma. In some embodiments, the individual has a pathological stage of "CIS+T1", where the individual has, simultaneously, one or more sites of bladder intraepithelial carcinoma and one or more sites of stage T1 papillary carcinoma.

[0089] "Papillary carcinoma" refers to bladder tumors that grow long, finger-like projections from the inner surface of the bladder toward the hollow center. Papillary tumors often grow toward the center of the bladder rather than into the deeper layers of the bladder. These tumors are called papillary carcinoma in situ. Very low grade (slow-growing) papillary carcinoma in situ is sometimes called low-grade papillary urothelial neoplasm (PUNLMP).

[0090] Stage "Ta" papillary carcinoma refers to papillary cancer found on the surface of the bladder lining. The cancer cells are clustered together and can often be easily removed. The cancer has not invaded the muscle or connective tissue of the bladder wall. Ta papillary carcinoma is also called noninvasive papillary urothelial carcinoma, or stage 0a bladder cancer.

[0091] Stage "T1" papillary carcinoma refers to papillary carcinoma that has grown through the inner wall of the bladder into the lamina propria. It has not spread to the thick muscular layer of the bladder wall or to lymph nodes or other organs. In some embodiments, the cancer has invaded the subepithelial connective tissue.

[0092] In some embodiments, the individual does not have concomitant papillary carcinoma associated with CIS. In some embodiments, the individual does not have low-grade or high-grade papillary carcinoma. In some embodiments, the individual does not have high-grade papillary carcinoma. In some embodiments, the individual does not have concomitant low-grade stage Ta papillary carcinoma. In some embodiments, the individual does not have concomitant high-grade stage Ta papillary carcinoma. In some embodiments, the individual does not have concomitant low-grade stage T1 papillary carcinoma. In some embodiments, the individual does not have concomitant high-grade stage T1 papillary carcinoma. In some embodiments, the individual does not have concomitant stage Ta or T1 papillary carcinoma.

[0093] In some embodiments, the individual has papillary carcinoma. In some embodiments, the individual has low-grade or high-grade papillary carcinoma in conjunction with CIS. In some embodiments, the individual has high-grade papillary carcinoma. In some embodiments, the individual has low-grade stage Ta papillary carcinoma in conjunction with CIS. In some embodiments, the individual has high-grade stage Ta papillary carcinoma in conjunction with CIS. In some embodiments, the individual does not have low-grade stage T1 papillary carcinoma in conjunction with CIS. In some embodiments, the individual does not have high-grade stage T1 papillary carcinoma in conjunction with CIS. In some embodiments, the individual does not have stage Ta or T1 papillary carcinoma in conjunction with CIS.

[0094] In some embodiments, the individual has papillary carcinoma without CIS. In some embodiments, the individual has low-grade or high-grade papillary carcinoma. In some embodiments, the individual has high-grade papillary carcinoma. In some embodiments, the individual has low-grade stage Ta papillary carcinoma. In some embodiments, the individual has high-grade stage Ta papillary carcinoma. In some embodiments, the individual does not have low-grade stage T1 papillary carcinoma. In some embodiments, the individual does not have high-grade stage T1 papillary carcinoma. In some embodiments, the individual has stage Ta or T1 bladder cancer without CIS.

[0095] Bladder cancer can be detected and staged using any method known in the art, including, but not limited to, urine cytology, urethral cystoscopy (UCS), biopsy (e.g., transurethral resection of bladder tumor (TURBT)), biopsy (e.g., transnasal resection of bladder tumor, or "TURBT"), computed tomography (CT or CAT) (e.g., CT urography), magnetic resonance imaging (MRI, i.e., MR urography), positron emission tomography (PET or PET-CT), ultrasound (e.g., renal ultrasound), ureteroscopy, and radiography (e.g., chest radiography). A defining feature of bladder CIS is the fact that the consistency and adherence of CIS cells is reduced. As a result, there are more cells in the urine, which can be detected by urine cytology. In addition, bladder CIS is associated with an increased likelihood of exfoliated epithelium at the time of biopsy. Tumor cells from urine or biopsy samples can be examined using immunohistochemistry, ELISA, RT-PCR, or other methods suitable for detecting urinary markers for bladder CIS or papillary tumors. Fluorescence cystoscopy, for example, using porphyrin-based photosensitizers, may be particularly suitable for detecting CIS lesions.

[0096] Thus, in some embodiments, a method of treating an individual having only bladder intraepithelial carcinoma is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 1410 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0097] In some embodiments, a method of treating an individual with bladder CIS is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule, and the individual does not have concomitant papillary carcinoma at stage Ta or T1. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0098] In some embodiments, a method of treating an individual having bladder CIS (e.g., pure CIS) is provided, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered in an amount of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12Virus particles) once a week for about three weeks or about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0099] In some embodiments, a method of treating an individual having stage T1 bladder cancer with CIS is provided, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered in an amount of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 Virus particles) once a week for about three weeks or about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0100] In some embodiments, a method of treating an individual having stage Ta bladder cancer with CIS is provided, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered in an amount of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 121000 virions) once a week for about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0101] In some embodiments, a method of treating an individual having stage T1 bladder cancer without CIS is provided, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 1000 virions) once a week for about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0102] In some embodiments, a method of treating an individual having stage Ta bladder cancer without CIS is provided, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 121000 virions) once a week for about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0103] In some embodiments, the individual has early stage bladder cancer, non-metastatic bladder cancer, non-invasive bladder cancer, non-muscle invasive bladder cancer, primary bladder cancer, locally advanced bladder cancer (e.g., locally unresectable advanced bladder cancer), metastatic bladder cancer, or bladder cancer in remission. In some embodiments, the bladder cancer is resectable localized, unresectable localized, or unresectable. In some embodiments, the bladder cancer is a high-grade non-muscle invasive cancer that has been resistant to standard intravesical (intravesical) treatments. In some embodiments, the individual has a high-grade non-muscle invasive CIS that has been resistant to standard intravesical (intravesical) treatments. In some embodiments, the individual has a urothelial (i.e., transitional cell) carcinoma.

[0104] The methods provided herein can be used to treat individuals (e.g., humans) diagnosed with or suspected of having bladder cancer. In some embodiments, the individual has undergone tumor resection, such as TURBT or partial cystectomy. In some embodiments, the individual has previously undergone TURBT alone. In some embodiments, the individual has previously undergone TURBT with concurrent radiochemotherapy. In some embodiments, the individual has previously undergone repeat TURBT. In some embodiments, the individual has previously undergone TURBT at least about 1, 2, 3, 4 or more times. In some embodiments, the individual has previously undergone maximal TURBT. In some embodiments, the individual has not undergone tumor resection. According to the NCCN guidelines for bladder cancer, bladder preservation with concurrent radiochemotherapy with maximal TURBT is generally reserved for patients with small solitary tumors, negative lymph nodes, no intraepithelial carcinoma, no tumor-associated hydronephrosis, and good bladder function prior to treatment.

[0105] In some embodiments, the methods provided herein can be used to treat individuals with bladder cancer who have not undergone TURBT or partial cystectomy prior to treatment with CG0070. In some embodiments, the individual has stage Ta or T1 cancer. In some embodiments, the individual is considered to be unresectable or have residual disease. In some embodiments, the individual has an underlying medical condition that may not be amenable to resection.

[0106] Thus, in some embodiments, a method is provided for treating an individual having stage T1 or Ta bladder cancer that has not been resected or is considered unresectable, the method comprising administering CG0070 intravesically to the individual, wherein the CG0070 is administered at a dose of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 121000 virions) once a week for about six weeks. In some embodiments, the method further comprises administering to the individual a transduction enhancer (such as DDM) prior to administration of CG0070. In some embodiments, CG0070 is not administered in combination with an immune checkpoint modulator. In some embodiments, CG0070 is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy).

[0107] In some embodiments, the individual has refused a cystectomy, such as a partial cystectomy or a radical cystectomy. In some embodiments, the individual is not suitable for a cystectomy, such as a partial cystectomy or a radical cystectomy. In some embodiments, the individual has a low tolerance for a cystectomy. In some embodiments, the individual is medically inoperable. In some embodiments, the individual has previously completed a course of external beam radiation therapy and has significant residual disease. In some embodiments, the individual has previously undergone bladder conservation therapy. In some embodiments, the individual is an elderly patient. In some embodiments, the individual is unable to tolerate a cystectomy due to age. In some embodiments, the individual is at least about any of 60, 65, 70, 75, 80, 85, 90, 95 years of age or older. In some embodiments, the individual has renal deficiency or failure. According to the NCCN guidelines for bladder cancer, partial cystectomy is indicated for T2 muscle invasive disease with a solitary lesion in a location suitable for partial resection with adequate margins and absence of intraepithelial carcinoma as determined by random biopsy. Radical cystectomy or cystoprostatectomy is indicated for residual high-grade T1 and muscle invasive disease. Cystectomy is usually performed within 3 months of diagnosis if the patient is not undergoing treatment. Additionally, cystectomy is indicated for recurrent or persistent bladder cancer of stage Ta, T1, or CIS after TURBT or BCG treatment.

[0108] In some embodiments, the individual is not suitable for radical cystectomy according to National Comprehensive Cancer Network (NCCN) guidelines.For example, the individual may be unsuitable for curative therapy due to frailty.Before the method of the present invention, such an individual typically undergoes palliative radiotherapy without chemotherapy (10 times 3.5Gy / fraction, or 7 times 7Gy / fraction, TURBT, or no treatment).

[0109] In some embodiments, the individual is unable to tolerate a radical cystectomy based on American Society of Anesthesiology (ASA) guidelines, for example, an individual unable to tolerate a radical cystectomy may be considered medically unfit for surgery requiring general or epidural anesthesia.

[0110] In other embodiments, an individual may not have a post-operative care system or caregivers as determined by the Comprehensive Geriatric Assessment provided by the American Society of Anesthesiologists. Under these guidelines, an individual is considered frail if they are unable to perform normal, independent activities of daily living and exhibit severe nutritional abnormalities, cognitive impairment, or comorbidities of cumulative illness rating scale for geriatrics (CISR-G) grades 3-4.

[0111] Thus, in some embodiments, a method of treating an individual with bladder cancer is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule, and the individual has refused or is unsuitable for a cystectomy (e.g., radical cystectomy). In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0112] The method of the present invention also provides important and significant therapeutic benefits compared to standard treatment regimens that call for the removal of the bladder.The present invention also has the advantage of being useful as a bladder-preserving protocol for individuals who are suitable for cystectomy but do not choose to undergo cystectomy.The method of the present invention allows the bladder to be preserved after the onset of bladder cancer, resulting in a significant improvement in the quality of life of individuals compared to currently available treatments.

[0113] Thus, in some embodiments, a method of bladder preservation is provided herein, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to an individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule, and the individual has refused or is unsuitable for a cystectomy (e.g., radical cystectomy). In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0114] In some embodiments, the individual has been previously treated for bladder cancer (also referred to as "pre-treatment"). In some embodiments, the pre-treatment comprises one or more (1, 2, 3, 4, 5 or more) therapeutic modalities, including, but not limited to, surgery (e.g., transurethral resection of the bladder tumor, or partial cystectomy), intravesical treatment (e.g., BCG or intravesical chemotherapy), radiation therapy, chemotherapy, immunotherapy, and combinations thereof.

[0115] In some embodiments, the methods of the invention are particularly suited to individuals who have relapsed at least 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after BCG therapy. In some embodiments, the methods are particularly suited to individuals who have relapsed at least 9 months after receiving BCG therapy. In some embodiments, the individual has CIS.

[0116] In some embodiments, the individual has been previously treated with standard therapy for bladder cancer. In some embodiments, the individual has been previously treated with standard therapy for papillary carcinoma in situ. In some embodiments, the individual has been previously treated with standard therapy for bladder CIS. In some embodiments, the conventional standard therapy is intravesical therapy, such as intravesical chemotherapy or intravesical immunotherapy. In some embodiments, the previous standard therapy is treatment with mitomycin C. In some embodiments, the previous standard therapy is treatment with interferon (such as interferon alpha). In some embodiments, the previous standard therapy is treatment with platinum-based drugs. In some embodiments, the previous standard therapy is treatment with mitomycin and thiotepa. In some embodiments, the previous standard therapy is treatment with cisplatin, doxorubicin, gemcitabine, and valrubicin. In some embodiments, the previous standard therapy is BCG treatment.

[0117] In some embodiments, the individual has bladder CIS in remission, progressive bladder CIS, persistent bladder CIS, or recurrent bladder CIS. In some embodiments, the individual is resistant to treatment of bladder CIS with other agents (e.g., BCG or chemotherapy). In some embodiments, the individual initially responds to treatment of bladder CIS with other agents (e.g., BCG or chemotherapy), but progresses after treatment. In some embodiments, the individual has persistent or recurrent bladder CIS after undergoing TURBT. In some embodiments, the individual has persistent or recurrent bladder CIS after undergoing intravesical treatment, such as intravesical BCG or intravesical chemotherapy (e.g., mitomycin C). In some embodiments, the individual has persistent or recurrent bladder CIS after undergoing at least two consecutive cycles of intravesical treatment and / or at least two intravesical agents (e.g., intravesical BCG followed by intravesical mitomycin C). In some embodiments, the individual has persistent or recurrent bladder CIS after receiving a combination of TURBT and one or more intravesical therapies.

[0118] In some embodiments, the individual has recurrent bladder CIS (e.g., pure CIS) after prior treatment (prior standard treatment, such as intravesical BCG). For example, the individual may be initially responsive to prior treatment, but develop bladder CIS after about any of the following: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, or 60 months after discontinuing prior treatment.

[0119] In some embodiments, the individual has previously been treated with BCG. In some embodiments, the individual has previously been treated with at least one intravesical BCG treatment (e.g., at least about any of 1, 2, 3, 4, 5, 6, or more). In some embodiments, the intravesical BCG treatment comprises weekly BCG instillation into the bladder for at least about 4 weeks, e.g., at least about any of 5, 6, 7, 8, 9 weeks, or more. In some embodiments, the individual has previously been treated with an induction course of intravesical BCG treatment (e.g., about 4 to about 9, e.g., about 6 weeks of BCG administration). In some embodiments, the individual has previously been treated with an induction course and at least one maintenance course of intravesical BCG treatment. In some embodiments, the individual has previously been treated with an induction course of 6 to 233 k of BCG, followed by a maintenance 3-week instillation at any of about 3, 6, 12, 18, 24, 30, and 36 months. In some embodiments, the individual is on maintenance intravesical BCG therapy for at least about 3 months, such as at least about any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36 months or more. In some embodiments, the individual has been on BCG therapy within about any of 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month before receiving the viral therapy of the present application. In some embodiments, the individual has severe adverse effects after intravesical BCG therapy. In some embodiments, the individual has bacteriuria, persistent gross hematuria, persistent severe local symptoms, or systemic symptoms. In some embodiments, the individual is unable to tolerate intravesical BCG therapy even after lowering the dosage.

[0120] In some embodiments, the individual has failed BCG treatment. In some embodiments, the individual has failed BCG treatment within about any of 24, 22, 20, 18, 16, 14, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 months after the last dose of BCG. In some embodiments, the individual has failed an induction course of intravesical BCG treatment. In some embodiments, the individual has failed BCG treatment during or after a maintenance course.

[0121] In some embodiments, the individual does not respond to BCG treatment. In some embodiments, the individual had a partial response to BCG treatment. In some embodiments, the individual could not tolerate the adverse effects of BCG treatment. In some embodiments, the individual has disease recurrence after BCG treatment. In some embodiments, the individual has disease recurrence after about 24, 22, 20, 18, 16, 14, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month or less after the last dose of BCG (e.g., BCG induction or BCG maintenance). In some embodiments, the individual has disease progression despite receiving a maintenance course of BCG treatment. In some embodiments, the individual had bladder CIS before BCG treatment. In some embodiments, the individual did not have bladder CIS before BCG treatment. In some embodiments, the individual had a papillary tumor (e.g., stage Ta or T1) before BCG treatment. In some embodiments, the individual had both bladder CIS and papillary tumor (eg, stage Ta or T1) prior to BCG treatment.

[0122] Thus, in some embodiments, a method of treating BCG-non-responsive non-muscle invasive bladder cancer (e.g., BCG-non-responsive bladder CIS) in an individual is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual has only bladder CIS. In some embodiments, the individual has refused or is not suitable for a cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 week to about 6 weeks (e.g., 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., 6 weeks).

[0123] In some embodiments, a method of treating an individual with bladder CIS is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule, and the individual has disease recurrence following BCG treatment. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual has bladder CIS only. In some embodiments, the individual has refused or is not suitable for a cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0124] Endpoints In some embodiments, the method prevents the progression of bladder cancer in an individual, hi some embodiments, the method prevents the progression of non-muscle invasive bladder cancer to muscle invasive bladder cancer for at least about any of 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or more.

[0125] In some embodiments, a method of preventing disease progression in an individual with bladder cancer is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0126] In some embodiments, the method prevents the growth of or reduces the size of the bladder cancer, hi some embodiments, the size of the bladder cancer is reduced by at least about 10% (including, for example, at least about any of 20%, 30%, 40%, 60%, 70%, 80%, 90%, or 100%).

[0127] In some embodiments, a method of inhibiting tumor growth or reducing tumor size in an individual with bladder cancer is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0128] In some embodiments, the method results in disease remission (partial or complete) in the individual, hi some embodiments, the individual is in disease remission for at least about any of 2, 3, 4, 5, 6, 12, 24 months, or more.

[0129] In some embodiments, a method is provided for effecting disease remission (e.g., partial or complete remission) in an individual with bladder CIS (e.g., pure CIS), the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of about 1 to about 6 weeks (e.g., about 3 weeks, or about 6 weeks). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or about 6 weeks).

[0130] In some embodiments, the method inhibits tumor metastasis in an individual. In some embodiments, at least about 10% (e.g., including any percentage of at least about 20%, 30%, 40%, 60%, 70%, 80%, 90%, or 100%) metastasis is inhibited. In some embodiments, a method is provided for inhibiting metastasis to lymph nodes. In some embodiments, a method is provided for inhibiting metastasis to lungs. Metastasis can be assessed by any method known in the art, such as blood tests, bone scans, x-rays, CT scans, PET scans, and biopsies.

[0131] In some embodiments, a method of inhibiting tumor metastasis in an individual with bladder cancer is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual is unresponsive to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0132] In some embodiments, the method extends survival (e.g., disease-free survival, progression-free survival, or cystectomy-free survival) of an individual. In some embodiments, survival is extended by at least about 2, 3, 4, 5, 6, 12, 24 months, or more.

[0133] In some embodiments, a method is provided for extending survival (e.g., disease-free survival, progression-free survival, or cystectomy-free survival) of an individual with bladder cancer, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor-selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0134] In some embodiments, the method improves the quality of life of the individual. In some embodiments, the individual does not require a cystectomy (e.g., partial or radical cystectomy) for at least about any of 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or more after receiving the viral therapy of the present application. In some embodiments, the method of the present application delays or eliminates the need for a radical cystectomy in the individual, so that the individual does not suffer from the unwanted side effects of reconstructive surgery after radical cystectomy and enjoys an improved quality of life compared to other patients with bladder CIS.

[0135] In some embodiments, a method of improving the quality of life of an individual with bladder CIS (e.g., pure CIS) is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0136] In some embodiments, a method of treating bladder cancer in an individual without subjecting the individual to a cystectomy (e.g., radical cystectomy) is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for a cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0137] In some embodiments, a bladder preservation method for treating bladder CIS (e.g., pure CIS) in an individual is provided, the method comprising intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual is unresponsive to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the method further comprises administering a transduction enhancing agent (such as DDM) to the individual prior to administration of the oncolytic virus. In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 2000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 weeks, or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer.

[0138] Treatment regimen Intravesical administration of oncolytic virus provides a unique opportunity to expose intravesical tumors to oncolytic virus in a relatively simple and effective manner, while potentially reducing toxicity to other tissues. The appropriate dose of oncolytic virus depends on factors such as the nature of the oncolytic virus, the bladder intraepithelial carcinoma being treated, and the route of administration. As used herein, "particles" as it relates to oncolytic virus refers to a collective number of physical units of oncolytic virus. This number can be converted by infectivity assays to another number, such as plaque forming units (pfu) or international units, which means infectious titer units, as is well known in the art, or is equivalent to such another number. In some embodiments, the oncolytic virus is about 1×10 5 Particles, 1×10 6 Particles, 1×10 7 Particles, 1×10 8 Particles, 1×10 9 Particles, 1×10 10 particles, 2×10 10 Particles, 5×10 10 Particles, 1×10 11 particles, 2×10 11 Particles, 5×10 11 Particles, 1×10 12 particles, 2×10 12 Particles, 5×10 12 Particles, 1×10 13 particles, 2×10 13 Particles, 5×10 13 Particles, 1×10 14 particles, or 1×10 15 In some embodiments, the oncolytic virus is administered at a dose of about 1×10 5 Particles ~ approx. 1 x 10 6 Particles, approximately 1 x 10 6 Particles ~ approx. 1 x 10 7 Particles, approximately 1 x 10 7 Particles ~ approx. 1 x 10 8 Particles, approximately 1 x 10 8 Particles ~ approx. 1 x 10 9 Particles, approximately 1 x 10 9 Particles ~ approx. 1 x 10 10 Particles, approximately 1 x 10 10 Particles ~ approx. 1 x 1011 Particles, approximately 1 x 10 11 Particles ~ approx. 5 x 10 11 Particles, about 5 × 10 11 Particles ~ approx. 1 x 10 12 Particles, approximately 1 x 10 12 Particles ~ approx. 2 x 10 12 Particles, about 2 x 10 12 Particles ~ approx. 5 x 10 12 Particles, about 5 × 10 12 Particles ~ approx. 1 x 10 13 Particles, approximately 1 x 10 13 Particles ~ approx. 1 x 10 14 particles, or approximately 1×10 14 Particles ~ approx. 1 x 10 15 The particles are administered in a dose of any one of the following:

[0139] In some embodiments, the oncolytic virus is administered once a day. In some embodiments, the oncolytic virus is administered at least about any one of 1, 2, 3, 4, 5, 6, or 7 times per week (i.e., daily). In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered uninterruptedly once a week, once a week for 2 out of 3 weeks, once a week for 3 out of 4 weeks, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, once every 8 weeks, once a month, or every 2-12 months. In some embodiments, the interval between each administration is less than any one of about 6 months, 3 months, 1 month, 20 days, 15 days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the interval between each administration is greater than about any one of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, there is no break in the dosing schedule. In some embodiments, the interval between each administration is less than about 1 week.

[0140] The administration of the oncolytic virus can be for an extended period of time, such as from about 1 month to about 7 years. In some embodiments, the oncolytic virus is administered for a period of at least about any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months. In some embodiments, the oncolytic virus is administered for a period of at least 4 weeks or 6 weeks. In some embodiments, the oncolytic virus is administered once a week for 4 weeks every 3 months. In some embodiments, the oncolytic virus is administered once a week for 6 weeks every 3 months.

[0141] In some embodiments, each therapeutic course includes weekly administration of the oncolytic virus for about 3 weeks or about 6 weeks. In some embodiments, the interval between each therapeutic course is at least about 1, 2, 3, 4, 5, 6 months, or more. In some embodiments, the interval between each therapeutic course is about 6, 5, 4, 3, 2, 1 month, or less. In some embodiments, the therapeutic course is repeated at least once, such as at least about 1, 2, 3, 4, 5, 6, or more times. In some embodiments, an individual is treated with an initial course of oncolytic virus followed by a maintenance course. In some embodiments, the initial course and the maintenance course have the same dose and schedule. In some embodiments, the initial course and the maintenance course have different doses and / or schedules.

[0142] In some embodiments, the oncolytic virus is administered by injecting a solution through a catheter, in some embodiments, the total volume of solution used for intravesical installation is about any of 1 mL, 10 mL, 50 mL, 75 mL, 100 mL, 125 mL, 150 mL, 200 mL, 250 mL, 300 mL, 400 mL, or 500 mL. In some embodiments, the total volume of the solution used for intravesical instillation is any of about 1 mL to about 10 mL, about 10 mL to about 50 mL, about 50 mL to about 75 mL, about 75 mL to about 100 mL, about 100 mL to about 125 mL, about 75 mL to about 125 mL, about 100 mL to about 150 mL, about 150 mL to about 200 mL, about 200 mL to about 300 mL, about 300 mL to about 400 mL, about 400 mL to about 500 mL, about 50 mL to about 500 mL, about 50 mL to about 250 mL, or about 100 mL to about 250 mL.

[0143] In some embodiments, the oncolytic virus is about 1×10 8 ~Approx. 1×10 15 Particles (approx. 1×10 11 ~Approx. 1×10 14 Particles, for example, about 1 x 10 12 In some embodiments, the oncolytic virus is administered in a volume of about 50 to about 500 mL (e.g., about 100 mL) by infusion. In some embodiments, the oncolytic virus is administered in a volume of about 1×10 in about 50 mL. 12 It is administered in individual doses.

[0144] The solution of oncolytic virus may be retained in the bladder for a period of time prior to urination to achieve uniform distribution or sufficient exposure of the oncolytic virus among the bladder tumor cells. In some embodiments, the solution is retained in the individual's bladder for at least about 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, or more. In some embodiments, the solution is retained in the individual's bladder for about 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 10 minutes to about 20 minutes, about 20 minutes to about 30 minutes, about 30 minutes to about 45 minutes, about 45 minutes to about 50 minutes, about 50 minutes to about 1 hour, about 5 minutes to about 15 minutes, about 10 minutes to about 30 minutes, about 30 minutes to about 1 hour, or about 1 hour to about 2 hours. In some embodiments, the oncolytic virus (e.g., CG0070) is retained in the individual's bladder for about 45 minutes to about 50 minutes. In some embodiments, the efficiency of intravesical administration of an oncolytic virus is further enhanced by a pretreatment comprising intravesical administration of an effective amount of a transduction enhancing agent, such as DDM.

[0145] The methods described herein may further comprise administering a pretreatment composition intravesically to the individual prior to administration of the oncolytic virus. In some embodiments, the pretreatment composition comprises a transduction enhancer, such as N-dodecyl-β-D-maltoside (DDM). DDM is a non-ionic surfactant consisting of maltose derivatized with a single 12-carbon chain, which acts as a mild detergent and solubilizer. It has been used as a food additive and is known to enhance mucosal surface penetration in rodents, likely due to its effect on membrane-associated GAGs and tight junctions.

[0146] In some embodiments, a method of treating bladder cancer in an individual is provided, the method comprising: (1) intravesically administering a transduction enhancing agent (e.g., DDM) to the individual; and subsequently (2) intravesically administering an oncolytic virus (e.g., CG0070) to the individual, where the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual is unresponsive to BCG treatment or has disease recurrence following BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the oncolytic virus is administered at a concentration of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12 In some embodiments, the oncolytic virus is administered at a dose of 1000 to 15000 viral particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer. In some embodiments, the individual does not undergo resection prior to administration of the oncolytic virus. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone a transurethral resection of the bladder tumor (TURBT). In some embodiments, the bladder cancer is Ta or T1 unresectable bladder cancer. In some embodiments, a saline irrigation is performed prior to intravesically administering the transduction enhancing agent to the individual.

[0147] In some embodiments, a method of treating bladder cancer (e.g., CIS bladder cancer without TURBT or Ta or T1 grade bladder cancer) is provided, comprising: (1) administering intravesical saline irrigation; (2) administering a transduction enhancing agent (e.g., DDM) by intravesical irrigation; (3) administering the transduction enhancing agent (e.g., DDM) to the individual by intravesical instillation; and (4) administering an oncolytic virus (e.g., CG0070) intravesically to the individual, wherein the oncolytic virus comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for oncolytic virus replication and a heterologous gene encoding an immune-related molecule. In some embodiments, the oncolytic virus comprises a tumor selective promoter. In some embodiments, the oncolytic virus is not administered in combination with an immune checkpoint modulating agent. In some embodiments, the oncolytic virus is administered as a single therapeutic agent. In some embodiments, the individual does not respond to BCG treatment or has disease recurrence after BCG treatment. In some embodiments, the individual has refused or is not suitable for cystectomy (e.g., radical cystectomy). In some embodiments, the oncolytic virus is administered at a dose of about 1×10 8 ~Approx. 1×10 14 10 virus particles (e.g., about 1 × 10 12In some embodiments, the oncolytic virus is administered at a dose of 12 to 15 virus particles. In some embodiments, the oncolytic virus is administered once a week. In some embodiments, the oncolytic virus is administered for about 1 week to about 6 weeks (e.g., about 3 or 6 weeks). In some embodiments, the bladder cancer is CIS, CIS+Ta, CIS+T1, Ta or T1 grade bladder cancer. In some embodiments, the individual has Ta or T1 bladder cancer and has not undergone transurethral resection of the bladder tumor (TURBT). In some embodiments, the bladder cancer is Ta or T1 unresectable bladder cancer. In some embodiments, the individual does not undergo resection prior to administration of the oncolytic virus. In some embodiments, the transduction enhancing agent is retained in the bladder for 12 to 15 minutes during intravesical instillation. In some embodiments, a saline rinse is administered following intravesical instillation of the transduction enhancing agent and administration of the oncolytic virus.

[0148] The pretreatment composition is administered intravesically. In some embodiments, the pretreatment composition comprises a solution of a transduction enhancing agent (such as DDM). Suitable concentrations of the pretreatment composition (such as a DDM solution) include, but are not limited to, about any one of 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, or 5% of a transduction enhancing agent (such as DDM). In some embodiments, the pretreatment composition comprises any of about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.5% to about 1%, about 1% to about 2%, about 2% to about 3%, about 3% to about 4%, about 4% to about 5%, about 0.01% to about 1%, about 0.05% to about 2%, about 1% to about 5%, or about 0.1% to about 5% of a transduction enhancer (e.g., DDM).

[0149] Suitable doses of the pretreatment composition (such as DDM) include about 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 5 mg / kg, 10 mg / kg, 25 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, 250 mg / kg, 300 mg / kg, 400 mg / kg, 500 mg / kg, 0.1 mg / kg to 0.5 mg / kg, 0.5 mg / kg to 1 mg / kg, 1 mg / kg In some embodiments, the preferred dose of the pretreatment composition is about 0.1 g, 0.2 g, 0.5 g, 0.75 g, 1 g, 1.5 g, 2 g, 2.5 g, 5 g, or 10 g of the transduction enhancer (e.g., DDM).

[0150] In some embodiments, the pretreatment step is performed by contacting the luminal side of the bladder in the individual with a pretreatment composition prior to administration of the oncolytic virus. For example, the pretreatment composition may contain about 0.01% to about 0.5% (such as 0.05 to about 0.2%, for example about 0.1%) of a transduction enhancer (such as DDM). In some embodiments, the total volume of the pretreatment composition (such as DDM) is about 10 mL to about 1000 mL (such as about 10 mL to about 100 mL, about 100 mL to about 500 mL, or about 500 mL to about 1000 mL). In some embodiments, the pretreatment composition contains about 0.1% DDM. In some embodiments, a suitable dose of the pretreatment composition is any one of about 0.1 g, 0.2 g, 0.5 g, 0.75 g, 1 g, 1.5 g, 2 g, 2.5 g, 5 g, or 10 g of a transduction enhancer (such as DDM). In some embodiments, an effective amount of the pretreatment composition is about 1 g of DDM (eg, 100 mL of a 0.1% DDM solution).

[0151] In some embodiments, the pretreatment composition (such as DDM) is administered immediately prior to administration of the oncolytic virus (such as within 5 minutes prior to administration). In some embodiments, the pretreatment composition (such as DDM) is administered within about 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 90 minutes, 2 hours, 3 hours, or 4 hours prior to administration of the oncolytic virus. In some embodiments, the pretreatment composition (such as DDM) is administered within about 2 hours prior to administration of the oncolytic virus. In some embodiments, the pretreatment composition (such as DDM solution) is retained in the bladder for at least about any one of 5 minutes, 10 minutes, 15 minutes, or 20 minutes. In some embodiments, the pretreatment composition (such as DDM solution) is retained in the bladder for about 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 12 minutes to about 15 minutes, about 15 minutes to about 20 minutes, or about 10 minutes to about 20 minutes. In some embodiments, the pretreatment composition (eg, a DDM solution) is retained in the bladder for about 12 minutes to about 15 minutes.

[0152] In some embodiments, the pretreatment step is carried out by contacting the luminal side of the bladder in the individual with a pretreatment composition prior to administration of the oncolytic virus. In some embodiments, the method further comprises lavaging the luminal side of the bladder that is contacted with the pretreatment composition. In some embodiments, the method further comprises lavaging the luminal side of the bladder after contacting the bladder with the pretreatment composition prior to administration of the oncolytic virus.

[0153] In some embodiments, the pretreatment step comprises intravesical instillation of 75 mL or 100 mL of a pretreatment composition (eg, a DDM solution).

[0154] In some embodiments, the individual is a human individual. In some embodiments, the individual being treated for bladder intraepithelial carcinoma is identified as having one or more of the conditions described herein. Identification of the conditions described herein by a skilled physician is routine in the art (e.g., by blood tests, x-rays, ultrasound, CT scans, PET scans, PET / CT scans, MRI scans, PET / MRI scans, nuclear medicine radioisotope scans, endoscopy, biopsies, angiograms, CT angiograms, etc.), and these conditions may be suspected by the individual or others, for example, by tumor growth, bleeding, ulceration, pain, enlarged lymph nodes, cough, jaundice, swelling, weight loss, cachexia, sweating, anemia, paraneoplastic events, thrombosis, etc. In some embodiments, individuals are selected for any one of the treatment methods described herein based on any one or more of a number of risk factors and / or diagnostic techniques understood by those of skill in the art, including but not limited to genetic profiling, family history, medical history (e.g., manifestations of related conditions and history of viral infections), lifestyle or habits.

[0155] Oncolytic viruses The methods and compositions described herein relate to oncolytic viruses, e.g., oncolytic adenoviruses. Oncolytic viruses can be naturally occurring viruses or genetically engineered viruses, e.g., attenuated viruses, and / or viruses with additional favorable characteristics (e.g., preferential replication in cancer cells or encoding immune-related molecules).

[0156] Exemplary viruses suitable for use in the present invention include, but are not limited to, adenoviruses, such as H101 (ONCOCRINE®), CG-TG-102 (Ad5 / 3-D24-GM-CSF), and CG0070; herpes simplex viruses, such as Talimogene laherparapvec (T-VEC) and HSV-1716 (SEPREHVIR®); reoviruses, such as REOLYSIN®; vaccinia viruses, such as JX-594; Seneca Valley viruses, such as NTX-010 and SVV-001; Newcastle disease viruses, such as NDV-NS1 and GL-ONC1; polioviruses, such as PVS-RIPO; measles viruses, such as MV-NIS; coxsackie viruses, such as CAVATAK™; vesicular stomatitis viruses; Maraba and rhabdoviruses; parvoviruses and mumps viruses.

[0157] In some embodiments, the oncolytic virus is a wild-type oncolytic virus. In some embodiments, the oncolytic virus is genetically modified. In some embodiments, the oncolytic virus is attenuated (e.g., by multiple passages, inactivation, or genetic modification). In some embodiments, the oncolytic virus is propagation-prone. In some embodiments, the oncolytic virus replicates preferentially in cancer cells, such as cancer cells with a defect in the Rb pathway.

[0158] In some embodiments, the oncolytic virus (e.g., an oncolytic adenovirus) comprises a viral vector comprising a tumor cell specific promoter operably linked to a viral gene essential for oncolytic viral replication. In some embodiments, the tumor specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1, below. In some embodiments, the viral gene essential for oncolytic viral replication is selected from the group consisting of E1A, E1B, and E4.

[0159] In some embodiments, the oncolytic virus (e.g., an oncolytic adenovirus) comprises a viral vector comprising a tumor-selective promoter operably linked to a viral gene essential for oncolytic viral replication. In some embodiments, the tumor-selective promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1, below. In some embodiments, the viral gene essential for oncolytic viral replication is selected from the group consisting of E1A, E1B, and E4.

[0160] SEQ ID NO:1 (E2F-1 promoter)

[0161] In some embodiments, the oncolytic virus (e.g., oncolytic adenovirus) comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for oncolytic viral replication and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine) operably linked to the viral promoter. In some embodiments, the tumor-specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the viral gene essential for oncolytic viral replication is selected from the group consisting of E1A, E1B, and E4. In some embodiments, the viral promoter operably linked to a nucleic acid encoding an immune-related molecule is an E3 promoter. In some embodiments, the immune-related molecule is GM-CSF.

[0162] In some embodiments, the oncolytic virus (e.g., oncolytic adenovirus) comprises a viral vector comprising a tumor cell selective promoter operably linked to a viral gene essential for oncolytic viral replication and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine) operably linked to the viral promoter. In some embodiments, the tumor selective promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1. In some embodiments, the viral gene essential for oncolytic viral replication is selected from the group consisting of E1A, E1B, and E4. In some embodiments, the viral promoter operably linked to a nucleic acid encoding an immune-related molecule is an E3 promoter. In some embodiments, the immune-related molecule is GM-CSF.

[0163] In some embodiments, the oncolytic virus is an adenovirus serotype 5, in which the endogenous E1a promoter and the E3 19 kD coding region of the native adenovirus are replaced with the human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (e.g., a cytokine or chemokine, e.g., GM-CSF). In some embodiments, the tumor-specific promoter is a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO:1.

[0164] In some embodiments, the oncolytic virus is CG0070, an adenovirus serotype 5 with an E2F promoter in the E1a gene and GM-CSF expression in the E3 gene.

[0165] CG0070 is a conditionally replicating oncolytic adenovirus (serotype 5) designed to preferentially grow in and kill Rb pathway-deficient cancer cells. The vector is transcriptionally regulated by a promoter (e.g., E2F-1 promoter) that is upregulated in Rb pathway-deficient tumor cells. In approximately 85% of all cancers, one or more genes in the Rb pathway, such as the tumor suppressor Rb gene, are mutated. In addition to its restricted growth, CG0070 also encodes the human cytokine GM-CSF, which is selectively expressed in infected tumor cells and stimulates an immune response against uninfected distant (e.g., metastases) and local tumor lesions.

[0166] Genomic structure of oncolytic adenoviral vector CG0070 The gene structure is shown diagrammatically in Figure 1. The product of the adenoviral early E1A gene is essential for efficient expression of other regions of the adenoviral genome. CG0070 has been engineered to express the E1A gene under the control of the human E2F-1 promoter, which confers tumor specificity to the E1A gene product. A polyadenylation signal (PA) was inserted 5' of the E2F-1 promoter to protect against transcriptional read-through that activates E1A expression. CG0070 contains the entire wild-type E3 region except for the 19 kD coding region. Direct comparison of E3-containing and E3-deleted oncolytic adenoviral vectors demonstrated the superiority of the E3-containing vector in tumor spreading and efficacy. Instead of the 19 kD gene, CG0070 carries the cDNA for human GM-CSF under the control of the endogenous E3 promoter (E3P). Similarly, the E3 promoter is activated by E1A, so that finally, both viral replication and GM-CSF expression are under the control of the E2F-1 promoter. The remainder of the viral vector backbone, including E2, E4, the late protein region, and the inverted terminal repeats (ITRs), is identical to the wild-type Ad5 genome.

[0167] CG0070 is produced in HeLa-S3 cells and released from infected HeLa-S3 cells by detergent cell lysis. CG0070 is purified from the lysate by chromatography and then formulated in 5% sucrose, 10 mM Tris, 0.05% polysorbate-80, 1% glycine, 1 mM magnesium chloride, pH 7.8.

[0168] CG0070 is supplied as a sterile, slightly opalescent, frozen liquid in stoppered glass vials. The particle concentration per mL (vp / mL) will be listed on the Certificate of Analysis for each lot of CG0070.

[0169] CG0070 has additional potential antitumor activity because it carries the cDNA of human GM-CSF, a key cytokine for generating long-lasting antitumor immunity. Thus, CG0070 is a preferentially growing oncolytic vector with the potential to attack tumors by two mechanisms: direct cytotoxicity as a growing vector and induction of host immune responses. In vitro and in vivo studies have been conducted to clarify the tumor selectivity and antitumor activity and safety of CG0070. See, for example, US Patent Publication No. 20150190505, the entire contents of which are incorporated herein by reference. III. Pharmaceutical Compositions, Kits, and Articles of Manufacture

[0170] In another aspect, kits, unit doses, and articles of manufacture are provided that are useful for any one of the methods described herein.

[0171] In some embodiments, a pharmaceutical composition is provided that includes an oncolytic virus (e.g., CG0070) and a pharma- ceutical acceptable carrier, where the oncolytic virus includes a viral vector that includes a tumor cell-specific promoter operably linked to a viral gene essential for the replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. The pharmaceutical composition can be used to treat bladder CIS (e.g., pure CIS) by any one of the methods described herein.

[0172] Suitable pharmaceutical carriers include sterile water; saline, glucose; glucose in water or saline; condensates of castor oil and ethylene oxide containing about 30 to about 35 moles of ethylene oxide per mole of castor oil; liquid acids; lower alkanols; oils such as corn oil; peanut oil, sesame oil, etc., with emulsifiers such as mono- or di-glycerides of fatty acids, or phosphatides, e.g., lecithin; glycols; polyalkylene glycols; suspending agents, e.g., aqueous solvents in the presence of sodium carboxymethylcellulose; sodium alginate; poly(vinylpyrrolidone); and the like, alone or with a suitable dispersing agent such as lecithin; polyoxyethylene stearate; and the like. The carrier may contain auxiliary agents, such as preservative stabilizers, wetting agents, emulsifiers, etc., for use with penetration enhancers. The final form can be sterilized and can easily pass through an injection device such as a hollow needle. By appropriate selection of solvents or excipients, the appropriate viscosity can be obtained and maintained. Pharmaceutical carriers may include active or inactive excipients for drug delivery, including polymeric and non-polymeric. Other pharma- ceutically acceptable carriers and their formulations are described in standard formulation treatises, such as Remington's Pharmaceutical Sciences by EW Martin.

[0173] The pharmaceutical compositions described herein may include other agents, excipients, or stabilizers to enhance the properties of the composition. Examples of suitable excipients and diluents include, but are not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl- and propylhydroxybenzoic acid, talc, magnesium stearate, and mineral oil. In some embodiments, the pharmaceutical composition is formulated to have a pH range of about 4.5 to about 9.0, such as any pH range of about 5.0 to about 8.0, about 6.5 to about 7.5, or about 6.5 to about 7.0. In some embodiments, the pharmaceutical composition can also be made isotonic with blood by the addition of a suitable osmotic agent, such as glycerol.

[0174] In some embodiments, a kit for treating bladder intraepithelial carcinoma (e.g., pure CIS) in an individual is provided, the kit comprising an oncolytic virus (e.g., CG0070), wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for the replication of the oncolytic virus and a heterologous gene encoding an immune-related molecule. In some embodiments, the kit further comprises a pretreatment composition comprising a transduction enhancer, such as N-dodecyl-β-D-maltoside (DDM). In some embodiments, the kit further comprises a device, material, and / or instructions for performing any one of the above methods. The medical device for intravesical delivery may comprise a catheter, for example, Rusch 173430 Foley Catheter&BARD LUBRI-SIL Foley Catheter #70516SI.

[0175] The instructions for use of the oncolytic virus (e.g., CG0070) generally include information regarding the dosage, administration schedule, and route of administration for the intended treatment. The container may be a unit dose, bulk container (e.g., multi-dose package), or sub-unit dose. For example, a kit may be provided that includes a sufficient dose of the oncolytic virus disclosed herein to provide effective treatment of an individual for an extended period of time, such as any of 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, 12 months, or more. The kit may also include multiple unit doses of the oncolytic virus and instructions for use, packaged in a quantity sufficient for storage and use in pharmacies, such as hospital pharmacies and compounding pharmacies.

[0176] The kits of the present invention are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. The kits may optionally provide additional components, such as cushioning and interpretive information. Thus, the present application also provides articles of manufacture that include vials (such as sealed vials), bottles, jars, flexible packaging, and the like.

[0177] The article of manufacture may include a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, and the like. The container may be formed from a variety of materials, such as glass or plastic. In general, the container holds a composition effective for treating bladder CIS as described herein and may have a sterile access port. The label or package insert indicates that the composition is used to treat a particular condition in an individual. The label or package insert will further include instructions for administering the composition to an individual. Articles of manufacture and kits including the combination therapies described herein are also contemplated.

[0178] Package insert refers to instructions customarily included in commercial packaging of a therapeutic product that provide information regarding directions, usage, dosage, administration, contraindications, and / or warnings regarding the use of such therapeutic product. In some embodiments, the package insert indicates that the composition is used for treating bladder intraepithelial carcinoma (e.g., pure CIS).

[0179] In addition, the article of manufacture may further comprise a second container comprising a pharma- ceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution, and may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes. EXAMPLES

[0180] Working Example The following examples are intended to be purely illustrative of the present invention and therefore should not be considered as limiting the present invention in any manner. The following examples and detailed description are offered by way of illustration and not by way of limitation.

[0181] Example 1: Phase II clinical trial of CG0070 for the treatment of BCG-non-responsive non-muscle invasive bladder cancer (NMIBC) This example describes an open-label, single-arm, phase II, multicenter study for the safety and efficacy of intravesical CG0070 monotherapy in patients with non-muscle invasive bladder cancer who have failed BCG treatment and who have refused cystectomy. Most NMIBC (Cis, Cis with Ta and / or T1, high-grade Ta or T1 with frequent or uncontrolled recurrence) patients who fail intravesical BCG treatment (standard of care) usually have no option other than cystectomy. Cystectomy is a surgery associated primarily with morbidity, mortality, and quality of life issues. Morbidity and prolonged tedious medical care are relevant for the rest of the patient's life after cystectomy. Most patients with NMIBC stage do not show signs of disease progression or metastasis to the muscle layer, making surgery a very difficult decision. There is an unmet need for treatment options in this patient population.

[0182] CG0070 is a replication-selective oncolytic adenovirus that destroys bladder tumor cells through its deleted retinoblastoma (Rb) pathway. Previous reports of intravesical CG0070 have shown promising activity in patients with high-grade NMIBC who had previously failed to respond to BCG. However, limited case numbers have prevented analysis of efficacy, especially for pathological subsets. An interim analysis was performed 6 months after the first intravesical administration of CG0070 to investigate the efficacy of CG0070 monotherapy in patients with bladder carcinoma in situ (CIS) with or without concomitant papillary carcinoma at stage Ta or T1.

[0183] Study design In this study, each patient received 1 × 10 12 A dose of 1000 viral particles was administered once a week for 6 weeks. Patients who achieved a partial or complete response 6 months after the initial intravesical intervention were maintained with the same course of intravesical CG0070 once a week for 6 weeks. Patients were followed up every 3 months for 24 months. At the 6-month follow-up, patients underwent cystoscopy, urinary cytology, and biopsy. If no suspicious lesions were found, random biopsies of the bladder were obtained from patients. Biopsy samples were evaluated by pathologists at each institution and at a central site, and both biopsy results were used to determine treatment response.

[0184] The primary outcome of the study is a measure of durable complete response rate (DCR) at 18 months from the date of first intravesical administration of CG0070. DCR is defined as the proportion of patients with a durable complete response that persists for at least 6 months (first interim analysis), at least 12 months from the date of first confirmed complete response, and at least 18 months from the date of first intravesical intervention. Secondary outcome measures included cystectomy-free survival at 18 months after initial intravesical treatment, complete response survival at 18 months after initial intravesical treatment, time to progression to muscle invasive disease at 18 months after initial intravesical treatment, overall survival at 18 and 24 months after initial intravesical treatment, change in PD-L1 status pre- and post-intervention at either cystectomy or biopsy up to 24 months, PD-L1 status of cancer cells and immune cells at the tumor site by IHC, proportion of organ-confined disease at the time of cystectomy, proportion of patients with no cancer cells in the regional lymph nodes at the time of cystectomy, complete response rate at 24 months after initial intravesical treatment, proportion of patients with a complete response lasting at least 12 months, proportion of patients with disease regression at 24 months after initial intravesical treatment, and proportion of patients with partial and / or complete response lasting less than 12 months.

[0185] To be eligible for the study, patients must meet all of the following criteria: 1. 18 years of age or older, including adults and seniors; 2. Patients must have pathologically confirmed high-grade disease (HG) of non-muscle invasive bladder cancer (NMIBC) as defined by the 2004 WHO classification system, 3. Patients must not have evidence of muscle-invasive disease, 4. Patients must be able to provide a sufficient biopsy sample to a central pathologist for histopathologically confirmed transitional cell (urothelial) carcinoma. Urothelial tumors with mixed histology (but with less than 50% variants) are eligible. 5. Patients must have received at least two prior intravesical treatments according to the recommended schedule. BCG must be one of the prior treatments administered. 6. Patients may have either failed BCG induction therapy within 6 months or have had successful BCG treatment followed by recurrence. The first standard intravesical BCG therapy must include at least 6 weeks of weekly treatment (acceptable range of instillations per course is 4-9). The second BCG therapy must include at least 2 weeks of treatment. 7. Patients have either Cis or Cis with Ta and / or T1 disease at or before enrollment. For patients with only Ta or T1 disease at enrollment and patients with no history of Cis, disease recurrence must have occurred within 12 months of the most recent intravesical treatment of any kind, or disease recurrence within 18 months of BCG maintenance, or disease recurrence within 24 months of BCG induction. 8. T1 patients must have signs in the muscles included in the most recent biopsy, or undergo a repeat TURBT prior to enrollment. 9. The investigator has clearly explained to the subject that delaying cystectomy may increase the risk of disease progression, which may result in serious and potentially fatal outcomes, and the patient has refused radical cystectomy in a signed, specific section of the informed consent. 10. Patients must be available for study entry within 10 weeks of their most recent diagnostic procedure, usually a diagnostic biopsy, transurethral resection of the bladder tumor (TURBT), or positive urinary cytology. 11.Eastern Cooperative Oncology Group (ECOG) performance status <2; 12. Not pregnant or breastfeeding, 13. Patients of childbearing potential must agree to use appropriate contraception. 14. Agree to study-specific informed consent and HIPAA mandates for release of individual health information; 15. Adequate baseline CBC, renal and liver function. Parameters include WBC>3000 / mm 3 , ANC>1,000 / mm3 , hemoglobin > 9.5 g / dL, and platelet count > 100,000 / mm 3 Adequate renal function, as described: serum creatinine <2.5 mg / dL; bilirubin, AST, and ALT, less than 2× upper limit of normal; PT / INR, PTT, and fibrinogen, within institutional acceptable limits; absolute lymphocyte count, ≥800 / μL, before first dose of CG0070.

[0186] Patients who meet any of the following exclusion criteria will be excluded from the study: 1. Prior systemic chemotherapy or radiation therapy for bladder cancer. Note: Prior immunotherapy or intravesical (administered into the bladder) chemotherapy for superficial disease is permitted. 2. History of anaphylactic reactions following exposure to humanized or human therapeutic monoclonal antibodies, hypersensitivity to GM-CSF or yeast-derived products, clinically significant allergic reactions, or any known hypersensitivity or prior reaction to any of the compounded excipients in the study drug, 3. Known infection with HIV, HBV or HCV, 4. Anticipated use of chemotherapy or radiation therapy during the study that is not specified in the research protocol; 5. Any underlying medical condition that, in the opinion of the investigator, may result in harm to the patient from administration of the study vector, obscuring the interpretation of an adverse event, or precluding adequate surgical resection. 6. Systemic therapy in any ongoing clinical trial within 28 days prior to enrollment, 7. Concomitant therapy with immunosuppressants or immunomodulators, including any systemic steroids (exceptions: inhaled or topically applied steroids, and acute and chronic standard dose NSAIDs are permitted). Short-term (i.e., ≤ 1 day) use of glucocorticoids is permitted to prevent reactions to the intravenous contrast agents used in CT scans. 8. Immunosuppressive therapy, including cyclosporine, antithymocyte globulin, or tacrolimus, within 3 months of study entry 9. Have received an investigational cancer vaccine treatment (e.g., dendritic cell therapy, heat shock vaccine) within the past year. 10. History of stage III or higher cancer, excluding urothelial carcinoma. Basal cell or squamous cell skin cancer must be adequately treated and subjects must be disease free at the time of enrollment. Subjects with a history of stage I or II cancer must be adequately treated and have been disease free for at least 2 years at the time of enrollment. 11. Progressive or persistent viral or bacterial infections, 12. All infections must be resolved and patients must remain afebrile without antibiotics for 7 days prior to study enrollment. 13. Urinary tract infections, especially including bladder infections, must be resolved prior to study enrollment, 14. Unwilling or unable to comply with the protocol or cooperate fully with the investigator and site personnel.

[0187] Interim analysis results Interim Analysis 1 At the interim analysis cutoff date in October 2016, 36 patients with residual high-grade Ta, T1, or carcinoma in situ (CIS)+ / -Ta / T1 had completed 6 months of follow-up in this phase II, single-arm, multicenter trial (NCT02365818). Inclusion criteria indicated at least two previous intravesical treatments for CIS, one of which was BCG treatment. Patients had either failed BCG induction therapy within 6 months or had been successfully treated with BCG and subsequently relapsed. Complete response (CR) at 6 months was defined as absence of disease on cytology, cystoscopy, and random biopsies.

[0188] As shown in Figure 2 and Table 1 below, of the 36 patients in this interim analysis, 18 had CIS, 4 had CIS+Ta, 3 had CIS+T1, 8 had Ta, and 3 had T1. The overall CR at 6 months was 44%. Of note, the CR at 6 months was 72.2% in pure CIS patients, 52% in CIS+ / -Ta / T1, 0% in CIS+Ta / T1, and 27% in pure Ta / T1. In CIS non-responders, 4 patients (22%) had persistent CIS at 6 months, and 1 patient (5.6%) progressed to CIS+T1. No CR was seen at 6 months in pure T1 or CIS+Ta / T1 patients. In both CIS+Ta / T1 patients (n=7), 5 had persistent Ta / T1+ / -CIS, while 2 had CIS in the 6-month biopsy. [Table 1]

[0189] All treatment-related adverse events (AEs) at 6 months were grade 1-3, with dysuria (47%), bladder spasms (44%), hematuria (36%), and urgency (33%) being the most common urinary events. AEs related to immunologic treatment included fatigue (11%) and chills (5.6%). Grade 3 treatment-related AEs included dye (5.6%) and hypotenuse (2.7%). There were no grade 4 / 5 treatment-related AEs.

[0190] Interim Analysis 2 The 6-month clinical response results from the interim analysis with a cutoff date of April 14, 2017 are summarized in Tables 2–5.

[0191] Patients without evidence of malignancy, either by pathology or urine cytology or by most recent pathology, were ineligible for the study. Patient samples were evaluated by the local and central pathologists to determine each patient's baseline stage. If the local and central pathologists disagreed but both deemed malignant, the stages determined by both the local and central pathologists were combined. If the local and central pathologists disagreed and only one deemed malignant, the stage determined by the pathologist who deemed malignant was used to determine the patient's malignancy. If patient samples were evaluated only by either the local or central pathologists, the single available result was used. If neither pathologist deemed malignant but the urine cytology was positive, the patient's stage was annotated as "unknown." If neither pathologist deemed malignant but the urine cytology result was missing or negative, the patient's stage was annotated as "negative." Pathology reports with "papillae" were entered as Ta+T1. Pathology reports with "high-grade UCC" were interpreted as "Ta" unless invasion into the lamina propria (T1) or muscularis (T2) was identified. If urine cytology was the only method of evaluation, the result was atypical or suspicious, and there was no pathology report, the stage was annotated as "negative." If urine cytology was the only method of evaluation and the second urine cytology result was negative, the time point was considered negative.

[0192] Patient clinical response was determined using the same staging rules as the baseline stage assessment above. Complete response (CR) patients included those assessed as "negative" at 6 months after CG0070 treatment. Progressive disease (PD) patients included those who were assessed as "negative" at a previous time point, but not currently assessed as "negative," and patients who progressed to T2 or greater disease. [Table 2] * Twenty-two of 67 patients were considered not evaluable (NE) at 6 months and assumed not to be in CR. [Table 3] [Table 4] [Table 5]

[0193] This phase II study shows that in patients with high-risk BCG-nonresponsive NMIBC, intravesical CG0070 produced an overall complete response rate of 46.7% at 6 months with acceptable levels of toxicity. Particularly high responses and limited progression were seen in patients with pure CIS. Ongoing follow-up of this study will be valuable in the BCG-nonresponsive NMIBC population.

[0194] final result A total of 67 patients with bladder cancer at stages CIS, CIS+Ta, CIS+T1, Ta, or T1 were treated with intravesical CG0070. Patients were evaluated for complete response at 6 and 12 months as described above. Table 7 shows the response rates to CG0070 in BCG non-responsive, BCG resistant, and post-BCG recurrent bladder cancer patients. BCG non-responsive patients show a 49% complete response rate at 6 months and a 30% complete response rate at 12 months. BCG resistant patients show a 56% complete response rate at 6 months and a 44% complete response rate at 12 months. Finally, post-BCG recurrent patients show a 35% complete response rate at 6 months and an 18% complete response rate at 12 months. This indicates that CG0070 is effective in treating BCG non-responsive, post-BCG recurrent, and BCG resistant patients. [Table 6] [Table 7]

[0195] The response of BCG-nonresponsive bladder cancer patients was also analyzed by cancer subtype, as summarized in Table 8. As shown in Table 8, patients with papillary carcinoma had slightly higher complete response rates than patients with CIS-containing cancer (62% vs. 46% CR at 6 months, 55% vs. 26% CR at 12 months). [Table 8]

[0196] The association between time to relapse and response to CG0700 therapy was also evaluated. Of the 14 subjects who relapsed after achieving CR, 64% (9 / 14) relapsed at 6 months after achieving CR at 3 months. If no relapse occurs after this time point, CR is considered to be sustainable beginning at month 9. No Ta / T1 patients relapsed after achieving CR.

[0197] In summary, this Phase II study of CG0700 demonstrated efficacy for the treatment of both CIS-containing and papillary bladder cancer. Patients with non-responsive, BCG-resistant, and post-BCG recurrent bladder cancer all demonstrated high response rates to CG0700, demonstrating efficacy in this undertreated patient population. The high efficacy of CG0700 therapy suggests that it may be a viable alternative to radical cystectomy, allowing bladder preservation in patients with non-muscle invasive bladder cancer, improving the quality of life for these patients.

[0198] Example 2: Phase III Clinical Trial of CG0070 in the Treatment of Patients with Bladder Cancer This study will demonstrate the efficacy of a treatment regimen with CG0070 that includes an induction and maintenance phase. Patients with bladder cancer will receive CG0070 once a week for 6 weeks in month 0 of the induction phase. At month 3, patients will be re-evaluated. Patients with persistent disease will continue induction therapy and receive CG0070 once a week for 6 weeks in month 3. Patients who demonstrate a complete response will begin the maintenance phase and receive CG0070 once a week for 3 weeks in month 3. At month 6, during the maintenance phase, all patients will receive CG0070 once a week for 3 weeks. At months 12 and 18, during the maintenance phase, patients will receive CG0070 once a week for 3 weeks. Patients will receive up to 21 doses of CG0700. Complete response rates will be evaluated.

[0199] Example 3: Treatment of patients with Ta or T1 non-muscle invasive bladder cancer who have not undergone TURBT This study will demonstrate the efficacy of treating patients with Ta or T1 non-muscle invasive bladder cancer who have not undergone transurethral resection of the bladder tumor (TURBT), for example in the neoadjuvant setting. Patients who have not undergone TURBT will be administered CG0070. Patients will include those who are not suitable for TURBT and those considered to have unresectable stage Ta or T1 bladder cancer. Complete response rates will be evaluated.

Claims

1. 1. A composition comprising an oncolytic virus for use in a method of treating bladder cancer or preserving the bladder in an individual having bladder cancer, the method comprising administering the composition intravesically to the individual during an induction phase and a maintenance phase, the induction phase comprising administering the composition at a dose of oncolytic virus of about 1 x 1012 virus particles to about 2 x 1013 virus particles once a week for six weeks, and the maintenance phase comprising administering the composition once a week for three weeks, the oncolytic virus being adenovirus serotype 5, the endogenous E1a promoter of the native adenovirus being replaced with a human E2F-1 promoter, and the endogenous E3 19 kD coding region of the native adenovirus being replaced with a nucleic acid encoding human GM-CSF.

2. 2. The composition of claim 1, wherein the composition is administered once a week for three weeks every three or six months during the maintenance phase.

3. 3. The composition of claim 1 or 2, wherein the induction phase comprises administering the composition to the individual at a dose of about 1 x 1012 viral particles once a week for six weeks.

4. The composition of any one of claims 1 to 3, wherein the start of the induction phase and the start of the maintenance phase are separated by about 3 months.

5. The composition of any one of claims 1 to 3, wherein the start of the induction phase and the start of the maintenance phase are separated by about 6 months.

6. 6. The composition of claim 1, wherein the induction phase comprises administering the composition to the individual at a dose of about 1 x 10 12 virus particles to about 2 x 10 13 virus particles of oncolytic virus once a week for 6 weeks at months 0 and 3 of the treatment regimen.

7. The composition of any one of claims 1 to 6, wherein the individual has non-muscle invasive bladder cancer.

8. The composition of any one of claims 1 to 7, wherein the individual has carcinoma in situ.

9. The composition of claim 8, wherein the individual does not have stage Ta or T1 papillary carcinoma.

10. The composition of any one of claims 1 to 7, wherein the individual has stage Ta or T1 bladder cancer.

11. The composition of claim 10 , wherein the individual does not have carcinoma in situ.

12. The composition of any one of claims 1 to 11, wherein the individual is non-responsive to BCG treatment.

13. The composition of any one of claims 1 to 11, wherein the individual has disease recurrence following BCG treatment.

14. The composition of any one of claims 1 to 11, wherein the individual has failed BCG treatment within about 6 months.

15. The composition of any one of claims 1 to 14, wherein the individual has not undergone a cystectomy.

16. The composition of claim 15, wherein the individual has refused or is not suitable for cystectomy.

17. The composition of any one of claims 1 to 16, wherein the oncolytic virus is CG0070.

18. The composition according to any one of claims 1 to 17, characterized in that the maintenance phase comprises administering the composition at a dose of about 1 x 1012 virus particles to about 2 x 1013 virus particles of oncolytic virus once a week for three weeks.

19. The composition of any one of claims 1 to 18, wherein the method further comprises intravesically administering to the individual a transduction enhancing agent prior to administration of the composition.

20. The composition according to any one of claims 1 to 19, characterized in that the composition is administered as a single therapeutic agent.

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