Methods for treating epithelioid cell tumors

Abstract

To provide methods and compositions for treating epithelioid cell tumors (such as a PEComa).SOLUTION: A method of treating an epithelioid tumor in an individual comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, there is provided a method of treating a PEComa in an individual in need thereof, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the limus drug is sirolimus. In some embodiments, the albumin is human albumin (such as human serum albumin). In some embodiments, the nanoparticles comprise sirolimus associated (e.g., coated) with albumin.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 62 / 186,252, filed June 29, 2015, the contents of which are incorporated herein by reference in their entirety.

[0002] The present invention relates to methods and compositions for the treatment of proliferative diseases belonging to the family of epithelioid cell tumors, such as perivascular epithelioid cell tumors (PEComas), by administering a composition comprising an mTOR inhibitor, such as a limus drug, and nanoparticles comprising albumin. [Background technology]

[0003] Perivascular epithelioid cell tumors (PEComas) are a family of rare mesenchymal neoplasms composed of histologically and immunohistochemically distinctive epithelioid cells. Examples of PEComa tumors include lymphangioleiomyomatosis (LAM), angiomyolipoma (AML), pulmonary clear cell 'sugar' tumors, and PEComa not otherwise specified (PEComa-NOS; a term referring to poorly characterized PEComas of various other anatomical origins). PEComas share a distinctive cell type, perivascular epithelioid cells, which are often organized as nests and sheets, or occasionally as spindle cells, in focal association with the vascular wall. See Hornick, JL et al., Histopathology, 48:75-82 (2006); Wildgruber, M. et al., World J Surg Oncol, 12:1-4 (2014).

[0004] Although most PEComas are benign, a subset of invasive PEComas exhibit malignant behavior, e.g., developing locally invasive or distant metastases. See Gennatas, C. et al., World J Surg Oncol, 10:1-4 (2012); Wagner, A. J. et al., J Clin Oncol, 28:835-840 (2010); Koenig, A. M. et al., J Med Case Reports, 3:1-5 (2009).

[0005] First-line treatment for PEComa is surgical resection. See Martignoni, G. et al., Virchows Arch, 452:119-132 (2008). Treatment regimens with chemotherapy and / or radiation therapy remain controversial. PEComas are rarely reported, hindering the use of clinical trials designed to investigate novel regimens for the treatment of PEComa. See Selvaggi, F. et al., BMC Surg, 11 (2011); Waters, P.S. et al., Int J Surg Case Reports, 3:89-91 (2012). Second-line treatments for inoperable, invasive, progressive, locally advanced, metastatic, or malignant PEComas currently do not exist. Thus, these subsets of PEComas remain very difficult to treat. The prognosis for patients within these patient subsets is poor, with median survival estimated at 12 to 17 months after diagnosis of progressive disease. Further, for example, the 5-year survival rate for metastatic uterine PEComa is approximately 16%. See Khaja, F. et al., Case Reports in Medicine, 2013:1-4 (2013). The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Selvaggi, F. et al., BMC Surg (2011) Vol. 11 [Non-patent document 2] Waters, PS et al., Int J Surg Case Reports (2012) 3:89-91 [Non-patent document 3] Khaja, F. et al., Case Reports in Medicine (2013) 2013:1-4 Summary of the Invention [Means for solving the problem]

[0007] In some embodiments, the present application provides a method of treating an epithelioid tumor in an individual, the method comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the epithelioid tumor is a perivascular epithelioid cell tumor (PEComa). In some embodiments, the PEComa is selected from the group consisting of "pale" clear cell tumor of the lung, PEComa not otherwise specified (PEComa-NOS), angiomyolipoma, and lymphangioleiomyomatosis. In some embodiments, the epithelioid tumor is malignant. In some embodiments, the epithelioid tumor is locally advanced. In some embodiments, the epithelioid tumor is metastatic.

[0008] In some embodiments according to any of the above methods, the mTOR inhibitor is a limus drug, such as sirolimus.

[0009] In some embodiments according to any of the above methods, the effective amount of the mTOR inhibitor in the nanoparticle composition is about 10 mg / m to about 100 mg / m (e.g., about 45-100 mg / m, about 75-100 mg / m, or about 45 mg / m, about 50 mg / m, about 75 mg / m, or about 100 mg / m). In some embodiments, the nanoparticle composition is administered weekly. In some embodiments, the nanoparticle composition is administered for two weeks every three weeks. In some embodiments, the nanoparticle composition is administered for three weeks out of four weeks. In some embodiments, the nanoparticle composition is administered on days 1 and 8 of a 21-day cycle. In some embodiments, the nanoparticle composition is administered on days 1, 8, and 15 of a 28-day cycle.

[0010] In some embodiments according to any of the above methods, the nanoparticle composition is administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, intraocularly, transdermally, orally, intraportally, intrahepatically, hepatic artery infusion, or by inhalation. In some embodiments, the nanoparticle composition is administered intravenously. In some embodiments, the nanoparticle composition is administered subcutaneously.

[0011] In some embodiments according to any of the above methods, the nanoparticles in the composition have an average diameter of about 150 nm or less, eg, about 120 nm or less.

[0012] In some embodiments according to any of the above methods, the mTOR inhibitor in the nanoparticle is associated with (eg, coated with) albumin.

[0013] In some embodiments according to any of the above methods, the individual is a human.

[0014] In some embodiments according to any of the above methods, the individual is selected for treatment based on levels (e.g., elevated levels) of melanocyte markers (e.g., including HMB45, MelanA, and microphthalmia transcription factor) and smooth muscle markers (e.g., including smooth muscle actin, pan-muscle actin, h-caldesmon, and calponin). In some embodiments, the levels of the melanocyte markers and smooth muscle markers are determined by immunohistochemistry.

[0015] In some embodiments of any of the above methods, this individual is selected for treatment based on the mutation status of the gene selected from the group consisting of TSC1, TSC2, TFE3, RHEB, MTOR, AKT, PIK3CA and PTEN.In some embodiments, for example, as determined by gene sequencing, if this individual has a mutation in this gene, this individual is selected for treatment.In some embodiments, this gene sequencing is based on the sequencing of circulating DNA or cell-free DNA in blood sample.In some embodiments, this gene sequencing is based on the sequencing of DNA in tumor sample.

[0016] In some embodiments according to any of the above methods, the individual is selected for treatment based on the phosphorylation status of a protein selected from the group consisting of AKT, S6, S6K, and 4EBP1. In some embodiments, if the protein in the individual is phosphorylated, the individual is selected for treatment. In some embodiments, if the protein in the individual is not phosphorylated, the individual is selected for treatment. In some embodiments, the phosphorylation status of the protein is determined by immunohistochemistry.

[0017] In some embodiments according to any of the above methods, the individual is selected for treatment based on the level of a proliferation marker or an apoptosis marker, for example, the proliferation marker Ki-67, or the apoptosis marker PARP or a fragment thereof. In some embodiments, the individual is selected for treatment based on the level of the proliferation marker and the apoptosis marker, for example, as determined by immunohistochemistry.

[0018] In some embodiments according to any of the above methods, the individual has not been previously treated with an mTOR inhibitor.

[0019] In some embodiments according to any of the above methods, the individual has been previously treated with chemotherapy, radiation, or surgery.

[0020] These and other aspects and advantages of the present invention will become apparent from the following detailed description and the appended claims. One, some, or all of the features of the various embodiments described herein may be combined to form other embodiments of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention provides methods and compositions for treating epithelioid cell tumors, such as perivascular epithelioid cell tumors (PEComas), in an individual in need thereof, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (hereinafter also referred to as an "mTOR nanoparticle composition") and albumin. In some embodiments, the composition comprises a limus drug and albumin (hereinafter also referred to as a "limus nanoparticle composition").

[0022] In some embodiments, a method of treating PEComa in an individual in need thereof is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the limus drug is sirolimus. In some embodiments, the albumin is human albumin (e.g., human serum albumin). In some embodiments, the nanoparticles comprise sirolimus associated with albumin (e.g., coated with albumin). In some embodiments, the average particle size of the nanoparticles in the nanoparticle composition is about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the composition comprises a nanoparticle formulation of sirolimus stabilized with albumin. In some embodiments, the composition is Nab-sirolimus.

[0023] In some embodiments, there is provided a method of treating PEComa in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug is associated with albumin (e.g., coated with albumin). In some embodiments, there is provided a method of treating PEComa in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticle composition has an average particle size of about 150 nm or less (e.g., less than about 120 nm). In some embodiments, there is provided a method of treating PEComa in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug is coated with albumin and the nanoparticle composition has an average particle size of about 150 nm or less (e.g., about 120 nm or less ... In some embodiments, a method of treating PEComa in an individual is provided, comprising administering to the individual an effective amount of Nab-sirolimus.

[0024] In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraportally. In some embodiments, the composition is administered intra-arterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatically. In some embodiments, the composition is administered by hepatic artery infusion. In some embodiments, the composition is administered intravascularly. In some embodiments, the composition is administered subcutaneously. In some embodiments, the composition is administered intrathecally. In some embodiments, the composition is administered intrapulmonary. In some embodiments, the composition is administered intramuscularly. In some embodiments, the composition is administered intratracheally. In some embodiments, the composition is administered intraocularly. In some embodiments, the composition is administered transdermally. In some embodiments, the composition is administered orally. In some embodiments, the composition is administered by inhalation.

[0025] PEComas that may be treated with the methods described herein include, but are not limited to, lymphangioleiomyomatosis (LAM), angiomyolipoma (AML), "pale" clear cell tumor of the lung, PEComa not otherwise specified (PEComa-NOS), and their malignant forms. In some embodiments, the PEComa is early-stage PEComa, non-metastatic PEComa, primary PEComa, advanced PEComa, locally advanced PEComa, metastatic PEComa, PEComa in remission, recurrent PEComa, PEComa in the add-on therapy setting, or PEComa in the neoadjuvant therapy setting. In some embodiments, the PEComa is resistant to treatment with a non-nanoparticle formulation of a chemotherapeutic agent (e.g., a non-nanoparticle formulation of a limus drug).

[0026] The methods described herein may be used for any one or more of the following purposes: alleviating one or more symptoms of PEComa, delaying the progression of PEComa, reducing tumor size in PEComa patients, inhibiting tumor growth of PEComa, extending overall survival, extending disease-free survival, extending the time to disease progression for PEComa, preventing or delaying PEComa tumor metastasis, reducing existing PEComa tumor metastasis, reducing the incidence or burden of existing PEComa tumor metastasis, and preventing recurrence of PEComa.

[0027] Also provided are compositions (such as pharmaceutical compositions), medicaments, kits, and unit doses useful in the methods described herein.

[0028] Also provided are methods of treating PEComa according to any one of the above methods, wherein the treatment is based on the expression or activity level or mutation status of one or more biomarkers, including but not limited to, mTOR pathway genes, including but not limited to, PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, RHEB, and TFE3, phosphoproteins, including but not limited to, p-AKT, p-S6, p-S6K, p-4EBP1, and p-SPARC, proliferation markers, including but not limited to, Ki-67, and apoptosis markers, including but not limited to, PARP.

[0029] definition As used herein, "treatment" or "treating" refers to an approach 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 the disease, reducing the extent of the disease, stabilizing the disease (e.g., preventing or delaying the progression of the disease), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying the recurrence of the disease, delaying or slowing the progression of the disease, improving the disease state, causing remission (partial or complete remission) of the disease, reducing the dose of one or more other medications required to treat the disease, delaying the progression of the disease, improving quality of life, and / or prolonging survival. "Treatment" also encompasses the alleviation of the pathological consequences of PEComa. The methods of the present invention contemplate any one or more of these aspects of treatment.

[0030] The term "individual" refers to a mammal, including, but not limited to, a human, cow, horse, cat, dog, rodent, or primate. In some embodiments, the individual is a human.

[0031] As used herein, an "at risk" individual is one who is at risk of developing PEComa. An "at risk" individual may or may not have detectable disease, and may or may not exhibit detectable disease prior to the treatment methods described herein. "At risk" means that an individual has one or more so-called risk factors, which are measurable parameters that correlate with the development of PEComa as described herein. Individuals with one or more of these risk factors have a higher likelihood of developing cancer than individuals without these risk factor(s).

[0032] The term "adjuvant setting" refers to a clinical situation in which an individual has a history of PEComa and is generally (but not necessarily) responsive to treatment, including, but not limited to, surgery (e.g., surgical resection), radiation therapy, and chemotherapy. However, because of their history of PEComa, these individuals are considered at risk for developing the disease. Treatment or administration in the "adjuvant setting" refers to a subsequent treatment regimen. The degree of risk (e.g., whether an individual in the adjuvant setting is considered "high risk" or "low risk") depends on multiple factors and most usually depends on the extent of the disease when first treated.

[0033] "Neoadjuvant setting" refers to the clinical situation in which the method is administered prior to primary / definitive treatment.

[0034] As used herein, "delaying" the development of PEComa means postponing, inhibiting, slowing, retarding, stabilizing, and / or delaying the onset of cancer. This delay can be of varying lengths of time, 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 can, in effect, encompass prevention, in that the individual does not develop the disease. A method for "delaying" the development of PEComa is one that reduces the likelihood of disease development within a given time frame and / or reduces the extent of disease within a given time frame compared to the absence of the method. Such comparisons are typically based on clinical trials using a statistically significant number of subjects. The development of PEComa can be detected using standard methods, including, but not limited to, computed tomography (CAT scan), magnetic resonance imaging (MRI), abdominal ultrasound, coagulation tests, arteriography, or biopsy. Development can refer to the progression of PEComa, including its early undetectable development, development, recurrence, and onset.

[0035] As used herein, the term "effective amount" refers to a quantity of a compound or composition sufficient to treat a specified disorder, condition, or disease, such as improving, alleviating, ameliorating, and / or delaying one or more of its symptoms. For PEComa, an effective amount includes an amount sufficient to shrink tumors and / or reduce the rate of tumor growth (such as inhibiting tumor growth), or prevent or delay other unwanted cell proliferation in PEComa. In some embodiments, an effective amount is an amount sufficient to delay the onset of PEComa. In some embodiments, an effective amount is an amount sufficient to prevent or delay recurrence. An effective amount can be administered in one or more administrations. In the case of PEComa, an effective amount of a drug or composition is capable of (i) reducing the number of epithelioid cells, (ii) reducing the size of the tumor, (iii) inhibiting, retarding, slowing, and preferably stopping to some extent the invasion of PEComa cancer cells into peripheral organs, (iv) inhibiting (e.g., slowing to some extent, and preferably stopping) tumor metastasis, (v) inhibiting tumor growth, (vi) preventing or delaying the onset and / or recurrence of tumors, and / or (vii) alleviating to some extent one or more symptoms associated with PEComa.

[0036] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" means a biologically or otherwise desirable material, e.g., a material that can be incorporated into a pharmaceutical composition administered to a patient without causing any significantly undesired biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. Pharmaceutically acceptable carriers or excipients preferably meet required toxicological and manufacturing testing standards and / or are included in the "Guide on Inactive Ingredients" established by the U.S. Food and Drug Administration.

[0037] The term "Nab" used herein refers to nanoparticle albumin binding.For example, Nab-sirolimus is a nanoparticle albumin-bound formulation of sirolimus.Nab-sirolimus is also known as Nab-rapamycin, which has been previously described, for example, see WO2008109163A1, WO2014151853, WO2008137148A2 and WO2012149451A1.

[0038] As used herein, the term "mutation status" refers to the state (e.g., including mutations) of a gene sequence compared to a wild-type or reference gene sequence.

[0039] It will be understood that aspects and embodiments of the invention described herein include "consisting of" and / or "consisting essentially of" aspects and embodiments.

[0040] Reference herein to an "approximate" value or parameter encompasses (and describes) the variation that surrounds the value or parameter itself. For example, a reference to "about X" encompasses the description of "X."

[0041] As used in this specification and the appended claims, the singular forms "a," "an," "or," and "the" include plural referents unless the context clearly dictates otherwise. How to Treat PEComa

[0042] The present invention provides methods for treating PECM in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides methods for treating PECM in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the average or mean diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0043] " mTOR inhibitor " used herein refers to the inhibitor of mTOR.mTOR is a serine / threonine specific protein kinase downstream of phosphatidylinositol 3-kinase (PI3K) / Akt (protein kinase B) pathway, and is an important regulator of cell survival, proliferation, stress and metabolism.MTOR pathway dysregulation has been found in many human cancers, and mTOR inhibition produces substantial inhibitory effect on tumor progression. mTOR inhibitors described herein include BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), rapamycin (also known as sirolimus or Rapamune), AZD8055, temsirolimus (also known as CCI-779 and Torisel), PI-103, Ku-0063794, INK 128, AZD2014, NVP-BGT226, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354, and eforolimus (also known as ridaforolimus or deforolimus).

[0044] In some embodiments, the mTOR inhibitor is a limus drug, including sirolimus and its analogs. Examples of limus drugs include, but are not limited to, temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the limus drug is selected from the group consisting of temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506).

[0045] In some embodiments, the PEComa is lymphangioleiomyomatosis. In some embodiments, the PEComa is an angiomyolipoma. In some embodiments, the PEComa is a pulmonary "pale" clear cell tumor. In some embodiments, the PEComa is an extrapulmonary "pale" clear cell 'sugar' tumor. In some embodiments, the PEComa is PEComa-NOS. In some embodiments, the PEComa is malignant.

[0046] In some embodiments, the PEComa is early-stage PEComa, non-metastatic PEComa, non-invasive PEComa, invasive PEComa, primary PEComa, progressive PEComa, locally advanced PEComa, metastatic PEComa, recurrent PEComa, or PEComa in remission. In some embodiments, the PEComa has been refractory to prior treatment. In some embodiments, the PEComa is resistant to treatment with a non-nanoparticulate formulation of a chemotherapeutic agent (e.g., a non-nanoparticulate formulation of an mTOR inhibitor, such as a limus drug).

[0047] In some embodiments, the PEComa is locally resectable (e.g., a tumor that can be completely removed by surgery), locally unresectable (e.g., a localized tumor may be unresectable because significant vasculature is involved), or unresectable (e.g., tumor features or patient conditions that prevent surgical removal).

[0048] In some embodiments, the PEComa is located in or closely associated with the kidney, bladder, prostate, uterus, ovary, vulva, vagina, lung, pancreas, liver, lymph nodes and / or skin.

[0049] In some embodiments, the PEComa is a stage I tumor (single tumor without vascular invasion), stage II tumor (single tumor with vascular invasion or multiple tumors 5 cm or less), stage III tumor (multiple tumors, any larger than 5 cm), stage IV tumor (tumor with direct invasion of adjacent organs other than the gallbladder or perforation of the visceral peritoneum), N1 tumor (regional lymph node metastasis), or M1 tumor (distant metastasis) according to the TNM classification. In some embodiments, the PEComa is a stage T1, T2, T3, or T4 PEComa according to the AJCC (American Joint Commission on Cancer) staging criteria. In some embodiments, the individual is at a clinical stage of Ta, Tis, T1, T2, T3a, T3b, or T4. In some embodiments, the individual is at a clinical stage of Tis, CIS, Ta, or T1.

[0050] In some embodiments, the PEComa is characterized by immunohistochemically positive expression of melanocyte and smooth muscle markers. In some embodiments, the melanocyte marker is selected from the group consisting of HMB45, MelanA, and microphthalmia transcription factor. In some embodiments, the smooth muscle marker is selected from the group consisting of smooth muscle actin, pan-muscle actin, h-caldesmon, and calponin. In some embodiments, PEComa characterization is made on a primary tumor biopsy. In some embodiments, PEComa characterization is made on a metastatic tumor biopsy. In some embodiments, an individual is diagnosed as having a PEComa by characterization of the tumor as having immunohistochemically positive expression of melanocyte and smooth muscle markers. In some embodiments, the melanocyte marker is selected from the group consisting of HMB45, MelanA, and microphthalmia transcription factor. In some embodiments, the smooth muscle marker is selected from the group consisting of smooth muscle actin, pan-muscle actin, h-caldesmon, and calponin. In some embodiments, the diagnosis is made on a primary tumor biopsy. In some embodiments, the diagnosis is made on a metastatic tumor biopsy.

[0051] In some embodiments, a method for treating localized resectable PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating localized resectable PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus. In some embodiments, the PEComa can be resected with multiple resections.

[0052] In some embodiments, a method for treating unresectable PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating unresectable PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0053] Thus, for example, in some embodiments, a method for treating unresectable PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating unresectable PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0054] Thus, for example, in some embodiments, a method for treating malignant PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating malignant PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0055] Thus, for example, in some embodiments, a method for treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating metastatic PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0056] Thus, for example, in some embodiments, a method for treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating recurrent PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0057] Thus, for example, in some embodiments, a method for treating locally advanced PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for treating locally advanced PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0058] Thus, for example, in some embodiments, a method is provided for stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the present invention provides a method for stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, the method comprises administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, the method comprises administering to an individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0059] The methods provided herein can be used to treat an individual (e.g., a human) diagnosed with or suspected of having a PEComa. In some embodiments, the individual is a human. In some embodiments, the individual is at least about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 years of age. In some embodiments, the individual is male. In some embodiments, the individual is female. In some embodiments, the individual has undergone tumor resection. In some embodiments, the individual has refused surgery. In some embodiments, the individual is medically inoperable.

[0060] In some embodiments, the individual is a human exhibiting one or more symptoms associated with PEComa. In some embodiments, the individual is in an early stage of PEComa. In some embodiments, the individual is in an advanced stage of PEComa. In some embodiments, the individual is genetically or otherwise predisposed (e.g., has risk factors) to developing PEComa. Individuals at risk for PEComa include, for example, individuals whose risk is determined by analysis of genetic or biochemical markers. These risk factors include, but are not limited to, age, sex, race, diet, previous disease history, presence of precursor disease, genetic considerations (e.g., tuberous sclerosis complex (TSC) mutation status), and environmental exposures. In some embodiments, individuals at risk for PEComa include, for example, individuals who have relatives who have experienced PEComa and individuals whose risk is determined by analysis of genetic or biochemical markers (e.g., expression and activity levels or mutation status). Biomarkers include, but are not limited to, MTOR pathway genes, including but not limited to, PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, RHEB, and TFE3, phosphoproteins, including but not limited to, p-AKT, p-S6, p-S6K, p-4EBP1, p-SPARC, proliferation markers, including but not limited to, Ki67, and apoptosis markers, including but not limited to, PARP or fragments thereof. In some embodiments, a blood sample or tumor biopsy from an individual is used to assess the biomarkers.

[0061] The methods provided herein may be practiced in the setting of add-on therapy. In some embodiments, the methods are practiced in a neoadjuvant setting, i.e., the methods may be practiced before primary / definitive therapy. In some embodiments, the methods are used to treat a previously treated individual. In some embodiments, the individual has not been previously treated. In some embodiments, the methods are used as first-line therapy. In some embodiments, the methods are used as second-line therapy. In some embodiments, the individual has not been previously treated with an mTOR inhibitor. In some embodiments, the individual has not been previously treated with a limus drug.

[0062] In some embodiments, a method for extending the time to disease progression in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the method extends the time to disease progression by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the composition includes Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0063] In some embodiments, a method for prolonging survival of an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the method prolongs survival of the individual by at least any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0064] In some embodiments, a method for alleviating one or more symptoms in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0065] In some embodiments, a method for improving quality of life in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0066] In some embodiments, the individual has been previously treated for PEComa (also referred to as "prior treatment"). In some embodiments, the individual is resistant to treatment of PEComa with other agents (e.g., non-nanoparticulate formulations of mTOR inhibitors). In some embodiments, the individual is initially responsive to treatment of PEComa with other agents but progresses after treatment. Prior treatment includes, but is not limited to, chemotherapy, radiation, and surgery. In some embodiments, the prior treatment was stopped for 28 days or more prior to initiation of the methods described herein. In some embodiments, the prior treatment is stopped for more than five half-lives of the prior treatment agent prior to initiation of the methods described herein.

[0067] In some embodiments, the individual has a recurrent PEComa after prior treatment. For example, the individual may initially respond to treatment with prior therapy, but develop a PEComa about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 24 months, about 36 months, about 48 months, or about 60 months after cessation of prior therapy.

[0068] In some embodiments, the individual is refractory to prior treatment.

[0069] In some embodiments, the individual has progressed on the prior treatment at the time of treatment, for example, the individual has progressed on the prior therapy within about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months.

[0070] In some embodiments, the individual is refractory to prior treatment.

[0071] In some embodiments, the individual is unsuitable for continuing the prior treatment, for example, due to failure to respond and / or due to toxicity.

[0072] In some embodiments, the individual is non-responsive to prior treatment.

[0073] In some embodiments, the individual is partially responsive or shows a less than desirable degree of responsiveness to prior treatment.

[0074] In some embodiments, a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise a limus drug associated with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus associated with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and wherein the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, there is provided a method of treating lymphangioleiomyomatosis in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus.In some embodiments, methods are provided for treating lymphangioleiomyomatosis in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the mTOR inhibitor (e.g., a limus drug) is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for 3 weeks out of 4 weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over 30 minutes.

[0075] In some embodiments, a method for treating angiomyolipoma in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method for treating angiomyolipoma in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method for treating angiomyolipoma in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, a method for treating angiomyolipoma in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating angiomyolipoma in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating angiomyolipoma in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), the weight ratio of human albumin to sirolimus in the composition being about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, a method for treating a patient with sirolimus in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nab-sirolimus. Methods for treating angiomyolipoma are provided. In some embodiments, methods for treating angiomyolipoma in an individual (e.g., a human) are provided, comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for 3 weeks out of 4 weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over 30 minutes.

[0076] In some embodiments, a method is provided for treating a "clear" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method is provided for treating a "clear" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method is provided for treating a "clear" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, a method is provided for treating a "clear" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating a "pale" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise a limus drug associated with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating a "pale" clear cell tumor of the lung in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus associated with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and wherein the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1).In some embodiments, methods are provided for treating pulmonary "pale" clear cell tumors in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus. In some embodiments, methods are provided for treating pulmonary "pale" clear cell tumors in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks out of every three weeks (e.g., days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for three weeks out of four weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over a 30-minute period.

[0077] In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles comprise the limus drug in association with albumin (e.g., coated with albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, wherein the nanoparticles comprise sirolimus in association with human albumin (e.g., coated with human albumin), and the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and wherein the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1).In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus. In some embodiments, methods are provided for treating PEComa-NOS in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks out of every three weeks (e.g., days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for three weeks out of four weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over a 30-minute period.

[0078] In some embodiments, a method of treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method of treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method of treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, a method of treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating metastatic PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating metastatic PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), wherein the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, a method of treating metastatic PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus.In some embodiments, methods are provided for treating metastatic PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for 3 weeks out of 4 weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over 30 minutes.

[0079] In some embodiments, a method of treating locally advanced PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method of treating locally advanced PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method of treating locally advanced PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, a method of treating locally advanced PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating locally advanced PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating locally advanced PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), the weight ratio of human albumin to sirolimus in the composition being about 9:1 or less (e.g., about 9:1 or about 8:1).In some embodiments, methods are provided for treating locally advanced PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus. In some embodiments, methods are provided for treating locally advanced PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks out of every three weeks (e.g., days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for three weeks out of four weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over a 30-minute period.

[0080] In some embodiments, a method of treating progressive malignant PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method of treating progressive malignant PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method of treating progressive malignant PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, a method of treating progressive malignant PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating progressive malignant PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating progressive malignant PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), the weight ratio of human albumin to sirolimus in the composition being about 9:1 or less (e.g., about 9:1 or about 8:1).In some embodiments, methods are provided for treating progressive malignant PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus. In some embodiments, methods are provided for treating progressive malignant PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks out of every three weeks (e.g., days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for three weeks out of four weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over a 30-minute period.

[0081] In some embodiments, a method of treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a method of treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, a method of treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with albumin (e.g., coated with albumin). In some embodiments, a method of treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating recurrent PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of treating recurrent PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), the weight ratio of human albumin to sirolimus in the composition being about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, a method of treating recurrent PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus.In some embodiments, methods are provided for treating recurrent PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for 3 weeks out of 4 weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over 30 minutes.

[0082] In some embodiments, methods are provided for stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, methods are provided for stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, methods are provided for stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, wherein the limus drug in the nanoparticles is associated with (e.g., coated with) albumin. In some embodiments, there is provided a method of stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less). In some embodiments, there is provided a method of stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin, the nanoparticles comprising a limus drug associated with albumin (e.g., coated with albumin), the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less).In some embodiments, there is provided a method of stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and human albumin, the nanoparticles comprising sirolimus associated with (e.g., coated with) human albumin, the nanoparticles having an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, there is provided a method of stabilizing (e.g., preventing or delaying the progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising Nab-sirolimus. In some embodiments, methods are provided for stabilizing (e.g., preventing or delaying progression of) PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of Nab-sirolimus. In some embodiments, the limus drug is administered at a dose of about 10 mg / m to about 150 mg / m, including, for example, about 45 mg / m to about 100 mg / m and about 75 mg / m to about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 100 mg / m. In some embodiments, the limus drug is administered at a dose of about 75 mg / m. In some embodiments, the limus drug is administered at a dose of about 56 mg / m. In some embodiments, the limus nanoparticle composition is administered weekly. In some embodiments, the limus nanoparticle composition is administered for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, the limus nanoparticle composition is administered for 3 weeks out of 4 weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the limus nanoparticle composition is administered by intravenous infusion over 30 minutes.

[0083] The methods described herein are useful for various aspects of treating PEComa. In some embodiments, a method is provided for inhibiting perivascular epitheloid cell proliferation (e.g., PEComa tumor growth) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, cell proliferation is inhibited by at least about 10% (e.g., at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticles in the composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug associated with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the limus nanoparticle composition is administered for three weeks out of four weeks (e.g., days 1, 8, and 15 of a 28-day cycle). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0084] In some embodiments, a method for preventing local recurrence (e.g., tumor recurrence after resection) in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, metastasis is inhibited by at least about 10% (e.g., at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0085] In some embodiments, a method for inhibiting PEComa tumor metastasis in an individual is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, metastasis is inhibited by at least about 10% (e.g., at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, a method for inhibiting metastasis to lymph nodes is provided. In some embodiments, a method for inhibiting metastasis to the lung is provided. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug associated with (e.g., coated with) albumin. In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0086] In some embodiments, a method is provided for reducing (e.g., radiating) existing PEComa tumor metastasis (e.g., lung metastasis or lymph node metastasis) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, metastasis is inhibited by at least about 10% (including, for example, at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, a method is provided for inhibiting metastasis to lymph nodes. In some embodiments, a method is provided for inhibiting metastasis to the lung. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug associated with (e.g., coated with) albumin. In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus in association with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0087] In some embodiments, a method is provided for reducing the incidence or burden of existing PEComa tumor metastasis (e.g., lung metastasis or lymph node metastasis) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticles in the composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0088] In some embodiments, a method for reducing the size of a PEComa in an individual is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the tumor size is reduced by at least about 10% (including, for example, at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0089] In some embodiments, a method for extending the time to disease progression in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the method extends the time to disease progression by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0090] In some embodiments, a method for extending survival of an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the method extends survival of the individual by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticle composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising a limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0091] In some embodiments, a method for alleviating one or more symptoms in an individual with PEComa is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticles in the composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0092] In some embodiments, a method is provided for inhibiting the progression of CIS (carcinoma in situ) lesions in an individual with PEComa, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the limus drug is sirolimus. In some embodiments, the limus drug in the nanoparticles of the composition is administered intravenously. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin). In some embodiments, the composition comprises nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising the limus drug in association with albumin (e.g., coated with albumin), the nanoparticles having an average diameter of about 150 nm or less. In some embodiments, the composition comprises nanoparticles comprising sirolimus associated with human albumin (e.g., coated with human albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of human albumin to sirolimus in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the average or mean diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the composition comprises Nab-sirolimus. In some embodiments, the composition is Nab-sirolimus.

[0093] In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 100 mg / m2. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 100 mg / m2, wherein the composition is administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 100 mg / m2, the composition being administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle), the dose being administered by intravenous infusion over 30 minutes. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 100 mg / m2, the composition being administered weekly for three weeks every four weeks (e.g., on days 1, 8, and 15 of a 28-day cycle).In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 100 mg / m2, wherein the composition is administered weekly for three weeks every four weeks (e.g., on days 1, 8, and 15 of a 28-day cycle), wherein the dose is administered by intravenous infusion over 30 minutes.

[0094] In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 75 mg / m. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 75 mg / m. The composition is administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 75 mg / m2, wherein the composition is administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle), wherein the dose is administered by intravenous infusion over 30 minutes. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered weekly for three weeks every four weeks (e.g., on days 1, 8, and 15 of a 28-day cycle).In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 75 mg / m2, wherein the composition is administered weekly for 3 weeks every 4 weeks (e.g., on days 1, 8, and 15 of a 28-day cycle), wherein the dose is administered by intravenous infusion over 30 minutes.

[0095] In some embodiments, a method of treating PEComa in an individual (e.g., a human) is provided, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 56 mg / m2. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 56 mg / m2, administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle). In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 56 mg / m2, administered weekly for two weeks every three weeks (e.g., on days 1 and 8 of a 21-day cycle), wherein the dose is administered by intravenous infusion over 30 minutes. In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 56 mg / m2, wherein the composition is administered weekly for three weeks every four weeks (e.g., on days 1, 8, and 15 of a 28-day cycle). In some embodiments, there is provided a method of treating PEComa in an individual (e.g., a human) comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising sirolimus and albumin (e.g., nanoparticles comprising sirolimus coated with albumin, such as Nab-sirolimus), wherein the composition is administered at a dose of 56 mg / m2, wherein the composition is administered weekly for three weeks every four weeks (e.g., on days 1, 8, and 15 of a 28-day cycle), wherein the dose is administered by intravenous infusion over 30 minutes.

[0096] Also provided are pharmaceutical compositions comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) for use in any of the methods for treating PEComa described herein. In some embodiments, these compositions comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin).

[0097] Use of biomarkers The present invention, in one aspect, provides methods of treating epithelioid cell tumors, such as PEComa, based on the expression or activity level or mutational status of one or more biomarkers.

[0098] In some embodiments, a method is provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a mutational status in a gene. In some embodiments, the gene is selected from the ONCOPANEL™ test (CLIA-certified). In some embodiments, the gene is an mTOR pathway gene. In some embodiments, the gene is an mTOR-related pathway gene. As used herein, "mTOR-related pathway gene" refers to a gene that encodes a molecule, such as a protein, in the mTOR signaling pathway or that interacts directly or indirectly with the mTOR signaling pathway. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutational status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutational status is identified in a sample from an individual via exome sequencing. In some embodiments, the mutational status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutational status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0099] In some embodiments, a method is provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain phosphorylation state of a protein associated with the mTOR signaling pathway. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation state is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0100] In some embodiments, there is provided a method of treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of a proliferation marker. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, there is provided a method of treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of an apoptosis marker. In some embodiments, the apoptosis marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptosis marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0101] In some embodiments, a method is provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the nanoparticles have an average diameter of about 150 nm or less, and the individual is selected for treatment based on the individual having a mutation status in a gene. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from mTOR pathway genes. In some embodiments, the gene is an mTOR-related pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutation status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutation status is identified in a sample from the individual via exome sequencing. In some embodiments, the mutation status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutation status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0102] In some embodiments, a method is provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the nanoparticles have an average diameter of about 150 nm or less, and the individual is selected for treatment based on the individual having a certain phosphorylation state of a protein associated with the mTOR signaling pathway. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation state is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0103] In some embodiments, a method is provided for treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the nanoparticles have an average diameter of about 150 nm or less, and the individual is selected for treatment based on the individual having a certain expression level of a proliferation marker. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, a method is provided for treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual, comprising administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of an apoptosis marker. In some embodiments, the apoptosis marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptotic marker is measured in a sample from the individual via immunohistochemistry, hi some embodiments, the sample is a tumor biopsy.

[0104] In some embodiments, methods are provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising: (a) assessing the mutational status of a gene in the individual; and (b) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on having a mutational status in the gene. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from mTOR pathway genes. In some embodiments, the gene is an mTOR-related pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutational status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutational status is identified in a sample from the individual via exome sequencing. In some embodiments, the mutation status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutation status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0105] In some embodiments, methods are provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising: (a) assessing the mutation status of a gene in the individual; and (b) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on having a certain phosphorylation state of a protein associated with the mTOR signaling pathway. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation state is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0106] In some embodiments, a method is provided for treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual having a certain expression level of a proliferation marker, the method comprising: (a) assessing the mutation status of a gene in the individual; and (b) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of a proliferation marker. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, a method is provided for treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual having a certain expression level of a proliferation marker, the method comprising: (a) assessing the mutation status of a gene in the individual; and (b) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of an apoptosis marker. In some embodiments, the apoptosis marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptosis marker is determined in a sample from an individual by immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0107] In some embodiments, methods are provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising: (a) assessing the mutation status of a gene in the individual; (b) selecting (e.g., identifying or recommending) a treatment for the individual based on the individual's mutation status in the gene; and (c) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from mTOR pathway genes. In some embodiments, the gene is an mTOR-related pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutation status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutation status is identified in a sample from the individual via exome sequencing. In some embodiments, the mutation status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutation status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0108] In some embodiments, methods are provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual, comprising: (a) assessing the mutation status of a gene in the individual; (b) selecting (e.g., identifying or recommending) a treatment for the individual based on the individual's phosphorylation status of the protein; and (c) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation status is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0109] In some embodiments, a method of treating PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) in an individual is provided, comprising: (a) assessing the mutation status of a gene in the individual; (b) selecting (e.g., identifying or recommending) a treatment for the individual based on the individual having an expression level of a proliferation marker; and (c) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, wherein the individual is selected for treatment based on the individual having a certain expression level of the proliferation marker. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, methods are provided for treating PEComa (e.g., malignant PEComa, e.g., metastatic or locally advanced PEComa) in an individual with a certain expression level of a proliferation marker, comprising: (a) assessing the mutation status of a gene in the individual; (b) selecting (e.g., identifying or recommending) a treatment for the individual based on the individual's expression level of the proliferation marker; and (c) administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the apoptosis marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptosis marker is determined in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0110] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the mutational status of a gene in the individual; and (b) selecting or recommending a treatment to the individual based on the individual's mutational status in the gene. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from an mTOR pathway gene. In some embodiments, the gene is an mTOR-associated pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutational status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutational status is identified in a sample from the individual via exome sequencing. In some embodiments, the mutation status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutation status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0111] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the phosphorylation status of a protein in the individual; and (b) selecting or recommending treatment to the individual based on the individual's phosphorylation status of a protein associated with the mTOR signaling pathway. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation status is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0112] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the expression level of a proliferation marker in the individual; and (b) selecting or recommending treatment to the individual based on the individual's expression level of the proliferation marker. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the expression level of an apoptotic marker in the individual; and (b) selecting (e.g., identifying or recommending) treatment to the individual based on the individual's expression level of a proliferation marker. In some embodiments, the apoptotic marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptotic marker is determined in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0113] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the mutational status of a gene in the individual; (b) selecting or recommending a treatment to the individual based on the individual's mutational status in the gene; and (c) administering to the selected individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from an mTOR pathway gene. In some embodiments, the gene is an mTOR-related pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutational status is identified in a sample from the individual via cell-free DNA sequencing. In some embodiments, the mutational status is identified in a sample from an individual via exome sequencing. In some embodiments, the mutational status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutational status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0114] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the phosphorylation status of a protein in the individual; (b) selecting or recommending a treatment to the individual based on the individual having phosphorylation on the protein; and (c) administering to the selected individual an effective amount of a composition comprising nanoparticles comprising the limus drug and albumin. In some embodiments, the protein is selected from the group consisting of AKT, S6, S6K, 4EBP1, and SPARC. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is phosphorylated at a specific amino acid site. In some embodiments, the protein is unphosphorylated. In some embodiments, the phosphorylation status is identified in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0115] In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the expression level of a proliferation marker in the individual; (b) selecting or recommending a treatment to the individual based on the individual's expression level of the proliferation marker; and (c) administering to the selected individual an effective amount of a composition comprising nanoparticles comprising a limus drug and albumin. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. In some embodiments, methods are provided for selecting (including identifying or recommending) a subject having a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) for treatment with a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising: (a) assessing the expression level of an apoptotic marker in the individual; (b) selecting or recommending treatment to the individual based on the individual's expression level of the apoptotic marker; and (c) administering to the selected individual an effective amount of a composition comprising nanoparticles comprising the limus drug and albumin. In some embodiments, the apoptotic marker is PARP or a fragment thereof. In some embodiments, the expression level of the apoptotic marker is determined in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0116] Also provided herein are methods for assessing whether an individual with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) is more or less likely to respond to a treatment based on the individual's mutation status in a gene, the treatment comprising a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising assessing the mutation status of the gene in the individual. In some embodiments, the method further comprises administering to the individual determined to be likely to respond to the treatment an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the presence of a mutation in the gene indicates that the individual is more likely to respond to the treatment, and the absence of a mutation in the gene indicates that the individual is less likely to respond to the treatment. In some embodiments, the gene is selected from the ONCOPANEL™ test. In some embodiments, the gene is selected from mTOR pathway genes. In some embodiments, the gene is an mTOR-related pathway gene. In some embodiments, the gene is selected from the group consisting of PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. In some embodiments, the gene is TFE3. In some embodiments, the mutation status is identified in a sample from an individual via cell-free DNA sequencing. In some embodiments, the mutation status is identified in a sample from an individual via exome sequencing. In some embodiments, the mutation status is identified in a sample from an individual via tumor biopsy mutation analysis. In some embodiments, the mutation status is identified in a sample from an individual via fluorescent in-situ hybridization. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a tumor biopsy. In some embodiments, the sample is obtained before the initiation of a treatment method described herein. In some embodiments, the sample is obtained after the initiation of a treatment method described herein.

[0117] Also provided herein are methods for assessing whether an individual with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) is more or less likely to respond to a treatment based on the individual having a certain phosphorylation state in a protein, where the treatment comprises a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, and the method comprises assessing the phosphorylation state of the protein in the individual. In some embodiments, the method further comprises administering to the individual determined to be likely to respond to the treatment an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, a phosphorylated protein indicates that the individual is more likely to respond to the treatment. In some embodiments, a non-phosphorylated protein indicates that the individual is less likely to respond to the treatment.

[0118] Also provided herein are methods for assessing whether an individual with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) is more or less likely to respond to a treatment based on the individual's expression level of a proliferation marker, the treatment comprising a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, the method comprising assessing the expression level of the proliferation marker in the individual. In some embodiments, the method further comprises administering to the individual determined to be likely to respond to the treatment an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the expression level of the proliferation marker indicates that the individual is more likely to respond to the treatment. In some embodiments, the proliferation marker is Ki-67. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy. Further provided herein are methods for assessing whether an individual with a PEComa (e.g., a malignant PEComa, e.g., a metastatic or locally advanced PEComa) is more or less likely to respond to a treatment based on the individual's expression level of an apoptotic marker, where the treatment comprises a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin, and the method comprises assessing the expression level of the apoptotic marker in the individual. In some embodiments, the method further comprises administering to the individual determined to be likely to respond to the treatment an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin. In some embodiments, the expression level of the apoptotic marker indicates that the individual is less likely to respond to the treatment. In some embodiments, the apoptotic marker is PARP or a fragment thereof. In some embodiments, the expression level of the proliferation marker is measured in a sample from the individual via immunohistochemistry. In some embodiments, the sample is a tumor biopsy.

[0119] The genetic mutation status can be assessed from a variety of sample sources. In some embodiments, the sample is a blood sample. In some embodiments, cell-free DNA is isolated from the blood sample. In some embodiments, the sample is a tumor biopsy. The genetic mutation status can be assessed through a variety of methods well known to those skilled in the art. In some embodiments, the genetic mutation status is assessed using cell-free DNA sequencing. In some embodiments, the genetic mutation status is assessed using next-generation sequencing. In some embodiments, the genetic mutation status of a gene isolated from a blood sample is assessed using next-generation sequencing. In some embodiments, the genetic mutation status is assessed using exome sequencing. In some embodiments, the genetic mutation status is assessed using fluorescent in-situ hybridization analysis. In some embodiments, the genetic mutation status is assessed before the initiation of a treatment method described herein. In some embodiments, the genetic mutation status is assessed after the initiation of a treatment method described herein. In some embodiments, the genetic mutation status is assessed before and after the initiation of a treatment method described herein.

[0120] The phosphorylation state of a protein can be assessed from a variety of sample sources. In some embodiments, the sample is a tumor biopsy. The phosphorylation state of a protein can be assessed through a variety of methods well known to those of skill in the art. In some embodiments, the phosphorylation state is assessed using immunohistochemistry. The phosphorylation state of a protein can be site-specific. The phosphorylation state of a protein can be compared to a control sample. In some embodiments, the phosphorylation state is assessed before the initiation of a treatment method described herein. In some embodiments, the phosphorylation state is assessed after the initiation of a treatment method described herein. In some embodiments, the phosphorylation state is assessed before and after the initiation of a treatment method described herein. In some embodiments, the protein is phosphorylated. In some embodiments, the protein is unphosphorylated. In some embodiments, the site-specific amino acid is phosphorylated. In some embodiments, the site-specific amino acid is unphosphorylated.

[0121] The expression level of a protein (e.g., a proliferation or apoptosis marker) may be elevated or elevated compared to a control sample. In some embodiments, the level of a protein may be elevated or elevated compared to a control sample. In some embodiments, the level of a protein in an individual is compared to the level of the protein in a control sample. In some embodiments, the level of a protein in a subject is compared to the level of the protein in multiple control samples. In some embodiments, multiple control samples are used to generate a statistic used to classify the level of a protein in an individual with PEComa. The phosphorylation state of the protein may be compared to a control sample. In some embodiments, the expression level is assessed before the initiation of a treatment method described herein. In some embodiments, the expression level is assessed after the initiation of a treatment method described herein. In some embodiments, the expression level is assessed before and after the initiation of a treatment method described herein.

[0122] The classification or ranking (e.g., high or low) of protein levels can be determined by comparing them to a statistical distribution of control levels. In some embodiments, the classification or ranking is in comparison with a control sample obtained from the individual. In some embodiments, the protein levels are classified or ranked by comparing them to a statistical distribution of control levels. In some embodiments, the protein levels are classified or ranked by comparing them to levels from a control sample obtained from the subject.

[0123] The control sample may be obtained using the same source and method as the non-control sample. In some embodiments, the control sample is obtained from a different individual (e.g., an individual without PEComa and / or an individual sharing a similar ethnic, age, and gender identity). In some embodiments, where the sample is a tumor tissue sample, the control sample may be a non-cancerous sample from the same individual. In some embodiments, multiple control samples (e.g., from different individuals) are used to determine the range of biomarker levels in a particular tissue, organ, or cell population. In some embodiments, the control sample is a cultured tissue or cell that has been determined to be an appropriate control. In some embodiments, the control is a cell that does not express the biomarker. In some embodiments, a clinically accepted normal level in a standardized test is used as the control level to determine the biomarker level. In some embodiments, the reference level of the biomarker in a subject is classified as high, intermediate, or low according to a scoring system, such as an immunohistochemistry-based scoring system. In some embodiments, the reference level of a biomarker in a subject is classified as a low sample if the score is less than or equal to the overall median score.

[0124] In some embodiments, the biomarker level is determined by measuring the level of the biomarker in an individual and comparing it to a control or reference (e.g., the median level for a given patient population or the level of a second individual). For example, if the level of the biomarker for a single individual is determined to be above the median level for the patient population, the individual is determined to have a high level of the biomarker. Alternatively, if the level of the biomarker for a single individual is determined to be below the median level for the patient population, the individual is determined to have a low level of the biomarker. In some embodiments, the individual is compared to a second individual and / or patient population that is responsive to the treatment. In some embodiments, the individual is compared to a second individual and / or patient population that is not responsive to the treatment. In any of the embodiments herein, these levels may be determined by measuring the level of a nucleic acid encoding the biomarker. For example, if the level of mRNA encoding the biomarker for a single individual is determined to be above the median level for the patient population, the individual is determined to have a high level of mRNA encoding the biomarker. Alternatively, if the level of mRNA encoding a biomarker for a single individual is determined to be below the median level for a patient population, then that individual is determined to have a low level of mRNA encoding a biomarker.

[0125] In some embodiments, the reference level of a biomarker is determined by obtaining a statistical distribution of biomarker levels.

[0126] In some embodiments, bioinformatics methods are used to determine and classify the levels of biomarkers. A number of alternative bioinformatics approaches have been developed to evaluate gene set expression profiles using gene expression profiling data. Methods include, but are not limited to, those described in Segal, E. et al., Nat. Genet. 34:66-176 (2003); Segal, E. et al., Nat. Genet. 36:1090-1098 (2004); Barry, WT et al., Bioinformatics 21:1943-1949 (2005); Tian, ​​L. et al., Proc Nat'l Acad Sci USA 102:13544-13549 (2005); Novak BA and Jain A N. Bioinformatics 22:233-41 (2006); Maglietta R et al., Bioinformatics 23:2063-72 (2007); Bussemaker HJ, BMC Bioinformatics 8 Suppl 6:S6 (2007).

[0127] In some embodiments, the protein expression level is determined, for example, by immunohistochemistry. For example, criteria for low or high levels can be established based on the number of positively stained cells and / or the intensity of staining, for example, by using an antibody that specifically recognizes the biomarker protein. In some embodiments, the biomarker level is low if less than about 1%, less than about 5%, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, or less than about 50% of the cells have positive staining. In some embodiments, the biomarker level is low if the staining is 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% less intense than the positive control staining.

[0128] In some embodiments, the biomarker level is high when more than about 40%, more than about 45%, more than about 50%, more than about 55%, more than about 60%, more than about 65%, more than about 70%, more than about 75%, more than about 80%, more than about 85%, or more than about 90% of the cells have positive staining. In some embodiments, the biomarker level is high when the staining is of similar intensity to the positive control staining. In some embodiments, the biomarker level is high when the staining is 80%, 85%, or 90% of the intensity of the positive control staining.

[0129] In some embodiments, strong, moderate, and weak staining are calibrated levels of staining, where ranges are established and staining intensities are binned within the ranges. In some embodiments, strong staining is staining in an intensity range above the 75th percentile, moderate staining is staining in an intensity range between the 25th and 75th percentile, and weak staining is staining in an intensity range below the 25th percentile. In some aspects, one skilled in the art familiar with a particular staining technique will adjust the bin size and define the staining categories.

[0130] In some embodiments, the biomarkers are assessed from blood samples. In some embodiments, the biomarkers are assessed from cell-free DNA samples. In some embodiments, the biomarkers are assessed using next-generation sequencing. In some embodiments, the biomarkers are assessed from tumor biopsies. In some embodiments, the biomarkers are assessed using immunohistochemistry.

[0131] Further provided herein are methods of directing treatment for PEComa by delivering a sample to a diagnostic laboratory for determination of biomarker levels; providing a control sample with a known level of the biomarker; providing an antibody to the biomarker; subjecting the sample and the control sample to binding by the antibody; and / or detecting the relative amount of antibody binding, the levels in the sample being used to provide a conclusion that the patient should receive treatment by any one of the methods described herein.

[0132] Also provided herein is a method of directing the treatment of a disease, the method further comprising the steps of reviewing or analyzing data regarding the presence (or level) of a biomarker in a sample; and providing a conclusion to an individual, such as a health care provider or administrator, about the individual's likelihood or suitability for responding to treatment, the conclusion being based on the review or analysis of the data. In one aspect of the invention, the conclusion is transmission of data over a network.

[0133] The ONCOPANEL™ test can be used to investigate the exon DNA sequences and intron regions of cancer-related genes to detect genetic abnormalities, including somatic mutations, copy number variations, and structural rearrangements, in DNA from samples of various sources (e.g., tumor biopsies or blood samples), thereby providing a candidate list of genetic abnormalities that may be mTOR-activating abnormalities. In some embodiments, the mTOR-related genetic abnormality is a genetic abnormality or abnormal level (e.g., expression level or activity level) in a gene selected from the ONCOPANEL™ test. See, for example, Wagle N. et al., Cancer discovery 2.1 (2012): 82-93.

[0134] An exemplary version of the ONCOPANEL™ test includes 300 cancer genes and 113 introns across 35 genes. The 300 genes included in the exemplary ONCOPANEL™ test are: ABL1, AKT1, AKT2, AKT3, ALK, ALOX12B, APC, AR, ARAF, ARID1A, ARID1B, ARID2, ASXL1, ATM, ATRX, AURKA, AURKB, AXL, B2M, BAP1, BCL2, BCL2L1, BCL2L12, BCL6, BCOR, BCORL1, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1, BUB1B, CADM2, CARD11, CB L, CBLB, CCND1, CCND2, CCND3, CCNE1, CD274, CD58, CD79B, CDC73, CDH1, CDK1, CDK2, CDK4, CDK5, CDK6, CDK9, CDKN1A, CDKN1B, CDKN1C, CDKN2A, CDKN2B, CDKN2C, CEBPA, CHEK2, CIITA, CREBBP, CRKL, CRLF2, CRTC1, CRTC2, CSF1R, CSF3R, CTNNB1, CUX1, CYLD, DDB2, DDR2, DEPDC5, DICER1, DI S3, DMD, DNMT3A, EED, EGFR, EP300, EPHA3, EPHA5, EPHA7, ERBB2, ERBB3, ERBB4, ERCC2, ERCC3, ERCC4, ERCC5, ESR1, ETV1, ETV4, ETV5, ETV6, EWS R1, EXT1, EXT2, EZH2, FAM46C, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, FAS, FBXW7, FGFR1, FGFR2, FGFR3, FGFR4, FH, FKBP9, FLCN, FLT1, FL T3, FLT4, FUS, GATA3, GATA4, GATA6, GLI1, GLI2, GLI3, GNA11, GNAQ, GNAS, GNB2L1, GPC3, GSTM5, H3F3A, HNF1A, HRAS, ID3, IDH1, IDH2, IGF1R, I KZF1, IKZF3, INSIG1, JAK2, JAK3, KCNIP1, KDM5C, KDM6A, KDM6B, KDR, KEAP1, KIT, KRAS, LINC00894, LMO1, LMO2, LMO3, MAP2K1, MAP2K4, MAP3K1,MAPK1, MCL1, MDM2, MDM4, MECOM, MEF2B, MEN1, MET, MITF, MLH1, MLL(KMT2A) MLL2(KTM2D), MPL, MSH2, MSH6, MTOR, MUTYH, MYB, MYBL1, MYC, MYCL1(MYCL) 、MYCN、MYD88、NBN、NEGR1、NF1、NF2、NFE2L2、NFKBIA、NFKBIZ、NKX2-1、NOTC H1, NOTCH2, NPM1, NPRL2, NPRL3, NRAS, NTRK1, NTRK2, NTRK3, PALB2, PARK2 AX5, PBRM1, PDCD1LG2, PDGFRA, PDGFRB, PHF6, PHOX2B, PIK3C2B, PIK3CA, PI K3R1, PIM1, PMS1, PMS2, PNRC1, PRAME, PRDM1, PRF1, PRKAR1A, PRKCI, PRKCZ. PRKDC, PRPF40B, PRPF8, PSMD13, PTCH1, PTEN, PTK2, PTPN11, PTPRD, QKI, RA D21, RAF1, RARA, RB1, RBL2, RECQL4, REL, RET, RFWD2, RHEB, RHPN2, ROS1, RPL 26. RUNX1, SBDS, SDHA, SDHAF2, SDHB, SDHC, SDHD, SETBP1, SETD2, SF1, SF3B 1, SH2B3, SLITRK6, SMAD2, SMAD4, SMARCA4, SMARCB1, SMC1A, SMC3, SMO, SOCS 1, SOX2, SOX9, SQSTM1, SRC, SRSF2, STAG1, STAG2, STAT3, STAT6, STK11, SUF U, SUZ12, SYK, TCF3, TCF7L1, TCF7L2, TERC, TERT, TET2, TLR4, TNFAIP3, TP53 TSC1, TSC2, U2AF1, VHL, WRN, WT1, XPA, XPC, XPO1, ZNF217, ZNF708, ZRSR2 You can use the ONCOPANEL(wireless) license plate ABL1, AKT3, ALK, BCL2, BCL6 BRAF, CIITA, EGFR, ERG, ETV1, EWSR1, FGFR1, FGFR2, FGFR3, FUS, IGH, IGL. JAK2, MLL, MYC, NPM1, NTRK1, PAX5, PDGFRA, PDGFRB, PPARG, RAF1, RARA, RET.It is contemplated by the present application that mTOR-activating abnormalities (e.g., genetic abnormalities and abnormal levels) of any of the genes included in any embodiment or version of the ONCOPANEL™ test, including but not limited to the genes and intron regions listed above, can serve as a basis for selecting individuals for treatment with mTOR inhibitor nanoparticle compositions.

[0135] Dosage and Methods of Administering Nanoparticle Compositions The dose of mTOR nanoparticles (e.g., limus nanoparticle composition) administered to an individual (e.g., a human) can vary depending on the particular composition, the mode of administration, and the type of PEComa being treated. In some embodiments, the amount of the composition is effective to produce an objective response (e.g., a partial or complete response). In some embodiments, the amount of the mTOR nanoparticle composition (e.g., limus nanoparticle composition) is sufficient to produce a complete response in an individual. In some embodiments, the amount of the mTOR nanoparticle composition (e.g., limus nanoparticle composition) is sufficient to produce a partial response in an individual. In some embodiments, the amount of the mTOR nanoparticle composition (e.g., limus nanoparticle composition) administered (e.g., when administered alone) is sufficient to produce an overall response rate of greater than any of about 20%, about 30%, about 40%, about 50%, about 60%, or about 64% in a population of individuals treated with the mTOR nanoparticle composition (e.g., limus nanoparticle composition). An individual's response to treatment of the methods described herein can be determined based on, for example, RECIST levels, cystoscopy (with or without biopsy), biopsy, cytology, and CT imaging.

[0136] In some embodiments, the amount of the mTOR nanoparticle composition (eg, a limus nanoparticle composition) is sufficient to produce a negative biopsy in the individual.

[0137] In some embodiments, the amount of the composition is sufficient to extend the progression-free survival of the individual. In some embodiments, the amount of the composition is sufficient to extend the overall survival of the individual. In some embodiments, the amount of the composition (e.g., when administered alone) is sufficient to provide a clinical benefit in greater than about 50%, about 60%, about 70%, or about 77% of a population of individuals treated with the composition (e.g., a limus nanoparticle composition).

[0138] In some embodiments, the amount of the composition is sufficient to reduce tumor size, reduce the number of cancer cells, or slow tumor growth rate by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 100% compared to the corresponding tumor size or pancreatic tumor growth rate in the same subject before treatment, or compared to the corresponding activity in other subjects who have not received this treatment.Standard methods, such as in vitro assays using purified enzymes, cell-based assays, animal models, or human trials, can be used to measure the magnitude of this effect.

[0139] In some embodiments, the amount of mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the composition is below a level that induces toxic effects (i.e., effects above clinically acceptable levels of toxicity) or is at a level that allows potential side effects to be controlled or tolerated when the composition is administered to an individual.

[0140] In some embodiments, the amount of the composition is close to the maximum tolerated dose (MTD) of the composition according to the same dosing regimen, hi some embodiments, the amount of the composition is greater than any of about 80%, about 90%, about 95%, or about 98% of the MTD.

[0141] In some embodiments, the effective amount of the mTOR inhibitor (e.g., a limus drug) in the nanoparticle composition is at least 25 mg / m, 30 mg / m, 50 mg / m, 60 mg / m, 75 mg / m, 80 mg / m, 90 mg / m, 100 mg / m, 120 mg / m, 125 mg / m, 150 mg / m, 160 mg / m, 175 mg / m, 180 mg / m, 190 mg / m, 200 mg / m, 210 mg / m, 220 mg / m, 230 mg / m, 240 mg / m, 250 mg / m, 260 mg / m, 275 mg / m, 280 mg / m, 290 mg / m, 300 mg / m, 310 mg / m, 320 mg / m, 330 mg / m, 340 mg / m, 350 mg / m, 360 mg / m, 375 mg / m, 380 mg / m, 390 mg / m, 400 mg / m, 410 mg / m, 420 mg / m, 430 mg / m, 440 mg / m, 450 mg / m, 460 mg / m, 470 mg / m, 480 mg / m, 490 mg / m, 500 mg / m, 510 mg / m, 520 mg / m, 530 mg / m, 540 mg / m, 550 mg / m, 560 mg / m, 570 mg / m, 580 mg / m, 590 mg / m, 600 mg / m, 610 mg / m, 620 mg / m, 630 mg / m, 640 mg / m, 650 mg m2, 200 mg / m2, 210 mg / m2, 220 mg / m2, 250 mg / m2, 260 mg / m2, 300 mg / m2, 350 mg / m2, 400 mg / m2, 500 mg / m2, 540 mg / m2, 750 mg / m2, 1000 mg / m2, or 1080 mg / m2 of an mTOR inhibitor (e.g., Sirolis). In various embodiments, the composition contains less than approximately any of 350 mg / m2, 300 mg / m2, 250 mg / m2, 200 mg / m2, 150 mg / m2, 120 mg / m2, 100 mg / m2, 90 mg / m2, 50 mg / m2, or 30 mg / m2 of an mTOR inhibitor (e.g., Sirolis). In some embodiments, the amount of mTOR inhibitor (e.g., Sirolis) per administration is less than approximately any of 25 mg / m2, 22 mg / m2, 20 mg / m2, 18 mg / m2, 15 mg / m2, 14 mg / m2, 13 mg / m2, 12 mg / m2, 11 mg / m2, 10 mg / m2, 9 mg / m2, 8 mg / m2, 7 mg / m2, 6 mg / m2, 5 mg / m2, 4 mg / m2, 3 mg / m2, 2 mg / m2, or 1 mg / m2. In some embodiments, an effective amount of an mTOR inhibitor (e.g., Siloris) in the composition is in any of the following ranges: about 1 to about 5 mg / m, about 5 to about 10 mg / m, about 10 to about 25 mg / m, about 25 to about 50 mg / m, about 50 to about 75 mg / m, about 75 to about 100 mg / m, about 100 to about 125 mg / m, about 125 to about 150 mg / m, about 150 to about 175 mg / m, about 175 to about 200 mg / m, about 200 to about 225 mg / m, about 225 to about 250 mg / m, about 250 to about 300 mg / m, about 300 to about 350 mg / m, or about 350 to about 400 mg / m.In some embodiments, the effective amount of the mTOR inhibitor (e.g., Siloris) in the composition is about 5 to about 300 mg / m2, e.g., about 100 to about 150 mg / m2, about 120 mg / m2, about 130 mg / m2, or about 140 mg / m2.

[0142] In some embodiments of any of the above aspects, the effective amount of the mTOR inhibitor (e.g., sirolimus) in the composition includes at least about any of 1 mg / kg, 2.5 mg / kg, 3.5 mg / kg, 5 mg / kg, 6.5 mg / kg, 7.5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, or 60 mg / kg. In various embodiments, the effective amount of the mTOR inhibitor (e.g., sirolimus) in the composition includes at least about any of 350 mg / kg, 300 mg / kg, 250 mg / kg, 200 mg / kg, 150 mg / kg, 100 mg / kg, 50 mg / kg, 25 mg / kg, 20 mg / kg, 10 mg / kg, 7.5 mg / kg, 6.5 mg / kg, 5 mg / kg, 3.5 mg / kg, 2.5 mg / kg, or 1 mg / kg of the mTOR inhibitor (e.g., sirolimus).

[0143] In some embodiments, the dosing frequency for administering the nanoparticle composition includes, but is not limited to, daily, every other day, every 3 days, every 4 days, every 5 days, every 6 days, every week without a break, 3 weeks out of 4 weeks, once every 3 weeks, once every 2 weeks, or 2 weeks out of 3 weeks. In some embodiments, the composition is administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 6 weeks, or about once every 8 weeks. In some embodiments, the composition is administered at least about 1, 2, 3, 4, 5, 6, or 7 times per week (i.e., daily). In some embodiments, the interval between each administration is less than about 6 months, 3 months, 1 month, 20 days, 15 days, 14 days, 13 days, 12 days, 11 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 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 administration schedule. In some embodiments, the interval between each administration is less than about 1 week.

[0144] In some embodiments, the dosing frequency is once every two days, ranging from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 times. In some embodiments, the dosing frequency is once every two days, ranging from 5 times. In some embodiments, the mTOR inhibitor (e.g., sirolimus) is administered for at least 10 days, with an interval of about 2 days or less between each administration, and the dose of the mTOR inhibitor (e.g., sirolimus) in each administration is about 0.25 mg / m to about 25 mg / m, or about 25 mg / m to about 50 mg / m, such as about 0.25 mg / m to about 250 mg / m, about 0.25 mg / m to about 150 mg / m, or about 0.25 mg / m to about 75 mg / m.

[0145] Administration of the composition can be extended for an extended period of time, such as from about 1 month to about 7 years, hi some embodiments, the composition is administered for at least about any of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months.

[0146] In some embodiments, the dose of the mTOR inhibitor (e.g., sirolimus) in the nanoparticle composition can range from 5 to 400 mg / m2 when given on a 3-week schedule, or from 5 to 250 mg / m2 (e.g., 80 mg / m2 to 150 mg / m2, e.g., 100 to 120 mg / m2) when given on a weekly schedule. For example, the amount of the mTOR inhibitor (e.g., sirolimus) can be about 60 to about 300 mg / m2 (e.g., about 260 mg / m2) on a 3-week schedule.

[0147] Other exemplary dosing schedules for administering nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) include, but are not limited to, 100 mg / m2 weekly without a break; 100 mg / m2 weekly for two weeks out of three weeks; 100 mg / m2 weekly for three weeks out of four weeks; 75 mg / m2 weekly without a break; 75 mg / m2 weekly for two weeks out of three weeks; 75 mg / m2 weekly for three weeks out of four weeks; 56 mg / m2 weekly without a break; 56 mg / m2 weekly for two weeks out of three weeks; 56 mg / m2 weekly for three weeks out of four weeks. The frequency of administration of the composition can be adjusted over the course of treatment based on the judgment of the administering physician.

[0148] In some embodiments, the individual is treated for at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 treatment cycles.

[0149] The compositions described herein allow the compositions to be infused into individuals for an infusion time of less than about 24 hours.For example, in some embodiments, the compositions are administered for an infusion time of less than about 24 hours, 12 hours, 8 hours, 5 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or 10 minutes.In some embodiments, the compositions are administered for an infusion time of about 30 minutes.

[0150] In some embodiments, exemplary doses of an mTOR inhibitor (in some embodiments, a limus drug, e.g., sirolimus) in a nanoparticle composition include, but are not limited to, any of about 50 mg / m2, 60 mg / m2, 75 mg / m2, 80 mg / m2, 90 mg / m2, 100 mg / m2, 120 mg / m2, 160 mg / m2, 175 mg / m2, 200 mg / m2, 210 mg / m2, 220 mg / m2, 260 mg / m2, and 300 mg / m2. For example, the dosage of an mTOR inhibitor in a nanoparticle composition can range from about 100 to 400 mg / m2 when given on a 3-week schedule, or from about 50 to 250 mg / m2 when given on a weekly schedule.

[0151] mTOR nanoparticle compositions (e.g., sirolimus nanoparticle compositions) can be administered to an individual (e.g., a human) via a variety of routes, including, for example, intravenous, intraarterial, intraperitoneal, intrapulmonary, oral, inhalation, intravesicular, intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, transmucosal, and transdermal routes. In some embodiments, sustained-release formulations of the compositions can be used. In some embodiments, the compositions are administered intravenously. In some embodiments, the compositions are administered intravesically. In some embodiments, the compositions are administered intraarterially. In some embodiments, the compositions are administered intraperitoneally.

[0152] In some embodiments in which the limus nanoparticle composition is administered intravesically, the dose of the mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the nanoparticle composition can be in the range of about 30 mg to about 400 mg in a volume of about 20 to about 150 ml, and can be retained in the bladder for, e.g., about 30 minutes to about 4 hours. In some embodiments, the nanoparticle composition is retained in the bladder for, e.g., about 30 minutes to about 1 hour, about 1 hour to about 2 hours, about 2 hours to about 3 hours, or about 3 hours to about 4 hours.

[0153] In some embodiments, the dosage of the mTOR inhibitor (e.g., a limus drug, such as sirolimus) is about 100 to about 400 mg, e.g., about 100 mg, about 200 mg, about 300 mg, or about 400 mg. In some embodiments, the limus drug is administered at about 100 mg weekly, about 200 mg weekly, about 300 mg weekly, about 100 mg twice weekly, or about 200 mg twice weekly. In some embodiments, this administration is further followed by a monthly maintenance dose (which may be the same as or different from the weekly dose).

[0154] In some embodiments in which the limus nanoparticle composition is administered intravenously, the dose of the mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the nanoparticle composition can be in the range of about 30 mg to about 400 mg. The compositions described herein allow for the infusion of the composition into an individual over an infusion time of less than about 24 hours. For example, in some embodiments, the composition is administered over an infusion period of less than about 24 hours, about 12 hours, about 8 hours, about 5 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, about 20 minutes, or about 10 minutes. In some embodiments, the composition is administered over an infusion period of about 30 minutes to about 40 minutes.

[0155] Nanoparticle Composition The nanoparticle compositions described herein include nanoparticles comprising (in various embodiments consisting essentially of) an mTOR inhibitor (e.g., a limus drug, such as sirolimus). The nanoparticles can further comprise albumin (e.g., human serum albumin or human albumin). Nanoparticles of poorly water-soluble drugs are disclosed, for example, in U.S. Patent Nos. 5,916,596; 6,506,405; 6,749,868; 6,537,579; and 7,820,788, each of which is incorporated by reference in its entirety, and also in U.S. Patent Application Publication Nos. 2006 / 0263434 and 2007 / 0082838; and PCT Patent Application No. WO08 / 137148.

[0156] In some embodiments, the composition comprises nanoparticles having an average or mean diameter of about 1000 nanometers (nm) or less, such as about or less than any of about 900, 800, 700, 600, 500, 400, 300, 200, 150, 120, and 100 nm. In some embodiments, the nanoparticles have an average or mean diameter of about 150 nm or less, such as about 120 nm or less. In some embodiments, the nanoparticles have an average or mean diameter of about 10 nm to about 150 nm. In some embodiments, the nanoparticles have an average or mean diameter of about 40 nm to about 120 nm. In some embodiments, the nanoparticles are sterile-filterable.

[0157] In some embodiments, the average diameter of the nanoparticles in the compositions described herein is about 150 nm or less, including, for example, about 140, 130, 120, 110, 100, 90, 80, 70, or 60 nm or less. In some embodiments, at least about 50% (e.g., at least about 60%, 70%, 80%, 90%, 95%, or 99%) of the nanoparticles in the composition are about 150 nm or less, including, for example, about 140, 130, 120, 110, 100, 90, 80, 70, or 60 nm or less. In some embodiments, at least about 50% (e.g., at least about 60%, 70%, 80%, 90%, 95%, or 99%) of the nanoparticles in the composition are within the range of about 20 nm to about 150 nm, including, for example, about 40 nm to about 120 nm.

[0158] In some embodiments, the albumin has sulfhydryl groups capable of forming disulfide bonds. In some embodiments, at least about 5% (e.g., at least about 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) of the albumin in the nanoparticle portion of the composition is cross-linked (e.g., cross-linked via one or more disulfide bonds).

[0159] In some embodiments, nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) are associated with (e.g., coated with) albumin (e.g., human albumin or human serum albumin). In some embodiments, the composition comprises an mTOR inhibitor (e.g., a limus drug, such as sirolimus) in both nanoparticle and non-nanoparticulate form (e.g., in the form of a solution or in the form of a soluble albumin / nanoparticle complex), and at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the mTOR inhibitor (e.g., a limus drug, such as sirolimus) in the composition is in nanoparticle form. In some embodiments, the mTOR inhibitor (e.g., a limus drug, such as sirolimus) in the nanoparticles constitutes about 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the nanoparticles. In some embodiments, the nanoparticles have a non-polymeric matrix. In some embodiments, the nanoparticles comprise a core of an mTOR inhibitor (eg, a limus drug, such as sirolimus) that is substantially free of polymeric material (eg, a polymer matrix).

[0160] In some embodiments, the composition comprises albumin in both the nanoparticle and non-nanoparticle portions of the composition, and at least about any of 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the albumin in the composition is in the non-nanoparticle portion of the composition.

[0161] In some embodiments, the weight ratio of albumin (e.g., human albumin or human serum albumin) to mTOR inhibitor in the nanoparticle composition is about 15:1 or less, e.g., about 10:1 or less, or about 18:1 or less. In some embodiments, the weight ratio of albumin (e.g., human albumin or human serum albumin) to mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the composition is within any of the following ranges: about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 13:1, about 4:1 to about 12:1, or about 5:1 to about 10:1. In some embodiments, the weight ratio of albumin to mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the nanoparticle portion of the composition is about 1:2, 1:3, 1:4, 1:5, 1:9, 1:10, 1:15 or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in the composition is any of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, or about 1:1 to about 1:1.

[0162] In some embodiments, the nanoparticle composition comprises one or more of the above characteristics.

[0163] The nanoparticles described herein may be present in a dry formulation (such as a lyophilized composition) or may be suspended in a biocompatible medium. Suitable biocompatible media include, but are not limited to, water, aqueous buffered media, saline, buffered saline, optionally buffered amino acid solutions, optionally buffered protein solutions, optionally buffered sugar solutions, optionally buffered vitamin solutions, optionally buffered synthetic polymer solutions, lipid-containing emulsions, and the like.

[0164] Human serum albumin (HSA) is a highly soluble globular protein with a molecular weight of 65K and consists of 585 amino acids. HSA is the most abundant protein in plasma, accounting for 70-80% of the colloid osmotic pressure of human plasma. The amino acid sequence of HSA contains a total of 17 disulfide bridges, one free thiol (Cys34), and a single tryptophan (Trp214). Intravenous use of HSA solutions has applications in the prevention and treatment of hypovolemic shock (see, e.g., Tullis, JAMA, vol. 237, pp. 355-360, 460-463 (1977) and Houser et al., Surgery, Gynecology and Obstetrics, vol. 150, pp. 811-816 (1980)), and in conjunction with exchange transfusion in the treatment of neonatal hyperbilirubinemia (see, e.g., Finlayson, Seminars in Thrombosis and Hemostasis, vol. 6, pp. 85-120 (1980)). Other albumins, such as bovine serum albumin, are also contemplated. The use of such non-human albumins may be appropriate in situations where these compositions are used in non-human mammals, such as, for example, veterinary medicine (including household pets and agricultural settings). Human serum albumin (HSA) has multiple hydrophobic binding sites (eight in total for fatty acids, which are endogenous ligands of HSA) that bind a diverse set of drugs, particularly neutral and negatively charged hydrophobic compounds (Goodman et al., "The Pharmacological Basis of Therapeutics," 9th ed., McGraw-Hill New York (1996)).Two high-affinity binding sites have been proposed in the IIA and IIIA subdomains of HSA, which are extremely elongated hydrophobic pockets with near-surface charged lysine and arginine residues that serve as attachment points for polar ligand features (e.g., Fehske et al., Biochem. Pharmcol. 30:687-92 (198a); Vorum, Dan. Med. Bull. 46:379-99 (1999); Kragh-Hansen, Dan. Med. Bull. 1441:131-40 (1990); Curry et al., Nat. Struct. Biol. 5:827-35 (1998); Sugio et al., Protein. Eng., 12, 439-46 (1999); He et al., Nature, 358, 209-15 (199b); and Carter et al., Adv. Protein. Chem., 45, 153-203 (1994). Sirolimus and propofol have been shown to bind to HSA (see, e.g., Paal et al., Eur. J. Biochem., Vol. 268 (No. 7), pp. 2187-91 (200a); Purcell et al., Biochim. Biophys. Acta, 1478(a), pp. 61-8 (2000); Altmayer et al., Arzneimittelforschung, Vol. 45, pp. 1053-6 (1995); and Garrido et al., Rev. Esp. Anestestiol. Reanim, Vol. 41, pp. 308-12 (1994)). Furthermore, docetaxel has been shown to bind to human plasma proteins (see, e.g., Urien et al., Invest. New Drugs, 14(b), pp. 147-51 (1996)).

[0165] The albumin (e.g., human albumin or human serum albumin) in the composition generally serves as a carrier for the mTOR inhibitor; i.e., the albumin in the composition makes the mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) more readily suspendable in or helps maintain this suspension in an aqueous medium compared to a composition that does not contain albumin. This can avoid the use of toxic solvents (or surfactants) to solubilize the mTOR inhibitor, which may reduce one or more side effects of administering an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) to an individual (e.g., a human). Thus, in some embodiments, the compositions described herein are substantially free (e.g., surfactant-free) of surfactants, such as Cremophor (or polyoxyethylated castor oil, including Cremophor EL® (BASF)). In some embodiments, the nanoparticle composition is substantially free (e.g., surfactant-free) of surfactants. A nanoparticle composition is "substantially free of Cremophor" or "substantially free of surfactant" if the amount of Cremophor or surfactant in the composition is not sufficient to cause one or more adverse reactions in an individual when the nanoparticle composition is administered to the individual. In some embodiments, the nanoparticle composition contains less than about 20%, 15%, 10%, 7.5%, 5%, 2.5%, or 1% of an organic solvent or surfactant. In some embodiments, the albumin is human albumin or human serum albumin. In some embodiments, the albumin is recombinant albumin.

[0166] The amount of albumin in the compositions described herein varies depending on other components in the composition.In some embodiments, the composition comprises albumin in an amount sufficient to stabilize the mTOR inhibitor (for example, limus drug, for example, sirolimus) in the form of aqueous suspension, for example, stable colloidal suspension (such as stable suspension of nanoparticles).In some embodiments, the amount of albumin is such that it reduces the sedimentation rate of the mTOR inhibitor (for example, limus drug, for example, sirolimus) in aqueous medium.For particle-containing compositions, the amount of albumin also depends on the size and density of nanoparticles of mTOR inhibitor.

[0167] The taxane is "stabilized" in an aqueous suspension if the mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) remains suspended in the aqueous medium (e.g., without visible precipitation or settling) for an extended period of time, such as at least about 0.1, 0.2, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, 60, or 72 hours. The suspension is generally, but not necessarily, suitable for administration to an individual (e.g., a human). The stability of the suspension is generally (but not necessarily) evaluated at storage temperatures, such as room temperature (e.g., 20-25°C) or refrigerated conditions (e.g., 4°C). For example, a suspension is stable at storage temperatures if, about 15 minutes after preparation of the suspension, the suspension exhibits no flocculation or particle aggregation visible to the naked eye or when viewed under an optical microscope at 1000x magnification. Stability can also be assessed under accelerated testing conditions, such as temperatures above about 40°C.

[0168] In some embodiments, albumin is present in an amount sufficient to stabilize the mTOR inhibitor (e.g., a limus drug, such as sirolimus) in the aqueous suspension at a particular concentration. For example, the concentration of the mTOR inhibitor (e.g., a limus drug, such as sirolimus) in the composition is about 0.1 to about 100 mg / ml, including, for example, about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 to about 8 mg / ml, about 4 to about 6 mg / ml, or about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor (e.g., a limus drug, such as sirolimus) is about any of 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, and 50 mg / ml. In some embodiments, the albumin is present in an amount that avoids the use of surfactants (such as Cremophor) such that the composition is free or substantially free of surfactants (such as Cremophor).

[0169] In some embodiments, the composition in liquid form comprises about 0.1% to about 50% (w / v) albumin (e.g., about 0.5% (w / v), about 5% (w / v), about 10% (w / v), about 15% (w / v), about 20% (w / v), about 30% (w / v), about 40% (w / v), or about 50% (w / v)). In some embodiments, the composition in liquid form comprises about 0.5% to about 5% (w / v) albumin.

[0170] In some embodiments, the weight ratio of albumin to mTOR inhibitor (e.g., a limus drug, such as sirolimus) in the nanoparticle composition is such that a sufficient amount of the mTOR inhibitor binds to or is transported by cells. Although the weight ratio of albumin to mTOR inhibitor must be optimized for different combinations of albumin and mTOR inhibitor, generally, the weight ratio (w / w) of albumin to mTOR inhibitor (e.g., a limus drug, such as sirolimus) is about 0.01:1 to about 100:1, about 0.02:1 to about 50:1, about 0.05:1 to about 20:1, about 0.1:1 to about 20:1, about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, or about 9:1. In some embodiments, the weight ratio of albumin to mTOR inhibitor is about any of 18:1 or less, 15:1 or less, 14:1 or less, 13:1 or less, 12:1 or less, 11:1 or less, 10:1 or less, 9:1 or less, 8:1 or less, 7:1 or less, 6:1 or less, 5:1 or less, 4:1 or less, and 3:1 or less. In some embodiments, the weight ratio of albumin (e.g., human albumin or human serum albumin) to mTOR inhibitor in the composition is any one of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, or about 1:1 to about 1:1.

[0171] In some embodiments, the albumin allows the composition to be administered to an individual (such as a human) without significant side effects. In some embodiments, the albumin (e.g., human serum albumin or human albumin) is in an amount effective to reduce one or more side effects of administering an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) to a human. The term "reducing one or more side effects" of administering an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) refers to the reduction, alleviation, elimination, or avoidance of one or more undesirable effects caused by the mTOR inhibitor, as well as side effects caused by the delivery vehicle used to deliver the mTOR inhibitor (e.g., a solvent that makes the limus drug suitable for injection). Such side effects include, for example, myelosuppression, neurotoxicity, hypersensitivity, inflammation, venous irritation, phlebitis, pain, skin irritation, peripheral neuropathy, neutropenic fever, anaphylactic reactions, venous thrombosis, extravasation, and combinations thereof. However, these side effects are exemplary only, and other side effects or combinations of side effects associated with mTOR inhibitors (eg, limus drugs, such as sirolimus) may also be reduced.

[0172] In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm).

[0173] In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, such as sirolimus) and albumin (e.g., human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm, and the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus and albumin (e.g., human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 100 nm), and the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or about 8:1.

[0174] In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in association with (e.g., coated with) albumin (e.g., human albumin or human serum albumin). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in association with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in association with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) in association with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus in association with (e.g., coated with) human albumin (e.g., human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm).

[0175] In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) associated with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) associated with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) associated with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm or less, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) associated with (e.g., coated with) albumin (e.g., human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus associated with (e.g., coated with) albumin (such as human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 100 nm), and the weight ratio of albumin to sirolimus in the composition is about 9:1 or about 8:1.

[0176] In some embodiments, the nanoparticle compositions described herein comprise an mTOR inhibitor (e.g., a ribozyme) stabilized with albumin (e.g., human albumin or human serum albumin). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus stabilized with human albumin (e.g., human serum albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 100 nm).

[0177] In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (e.g., human albumin or human serum albumin), wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (e.g., human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (such as human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm or less, and the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., a limus drug, e.g., sirolimus) stabilized with albumin (such as human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm, and the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the nanoparticle compositions described herein comprise nanoparticles comprising sirolimus stabilized with albumin (such as human albumin or human serum albumin), the nanoparticles having an average diameter of about 150 nm or less (e.g., about 100 nm), and the weight ratio of albumin to sirolimus in the composition is about 9:1 or about 8:1.

[0178] In some embodiments, the nanoparticle composition comprises Nab-sirolimus. In some embodiments, the nanoparticle composition is Nab-sirolimus. Nab-sirolimus is a formulation of sirolimus stabilized with human albumin USP and can be dispersed in directly injectable physiological solutions. The weight ratio of human albumin to sirolimus is about 8:1 to about 9:1. When dispersed in an appropriate aqueous medium, such as 0.9% sodium chloride injection or 5% dextrose injection, Nab-sirolimus forms a stable colloidal suspension of sirolimus. The average particle size of the nanoparticles in the colloidal suspension is about 100 nanometers. Because HSA is freely soluble in water, Nab-sirolimus can be reconstituted in a wide range of concentrations ranging from dilute (about 0.1 mg / ml sirolimus) to concentrated (about 20 mg / ml sirolimus), including, for example, about 2 mg / ml to about 8 mg / ml, or about 5 mg / ml.

[0179] The method of making nanoparticle composition is known in the art.For example, the nanoparticles containing mTOR inhibitor (for example, limus drug, for example, sirolimus) and albumin (such as human serum albumin or human albumin) can be prepared under high shear force conditions (for example, ultrasonic treatment, high-pressure homogenization, etc.).These methods are disclosed in, for example, United States Patent (USP) No. 5,916,596; United States Patent No. 6,506,405; United States Patent No. 6,749,868; United States Patent No. 6,537,579; and United States Patent No. 7,820,788, and also disclosed in United States Patent Application Publication No. 2007 / 0082838, United States Patent No. 2006 / 0263434 and PCT Application No. WO08 / 137148.

[0180] Briefly, mTOR inhibitor (for example, limus drug, for example, sirolimus) can be dissolved in organic solvent, and this solution can be added to albumin solution. This mixture is subjected to high-pressure homogenization. Then, the organic solvent can be removed by evaporation. The resulting dispersion can be further freeze-dried. Suitable organic solvents include, for example, ketones, esters, ethers, chlorinated solvents, and other solvents known in the art. For example, the organic solvent can be methylene chloride or chloroform / ethanol (for example, by the ratio of 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, or 9:1).

[0181] mTOR inhibitors In some embodiments, the methods described herein comprise the administration of nanoparticle compositions of mTOR inhibitors. As used herein, "mTOR inhibitor" refers to an inhibitor of mTOR. mTOR is a serine / threonine-specific protein kinase downstream of the phosphatidylinositol 3-kinase (PI3K) / Akt (protein kinase B) pathway and is an important regulator of cell survival, proliferation, stress, and metabolism. Dysregulation of the mTOR pathway has been found in many human cancers, and mTOR inhibition has a substantial inhibitory effect on tumor progression. In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor.

[0182] The mammalian target of rapamycin (mTOR), also known as the mechanistic target of rapamycin or FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1), is an atypical serine / threonine protein kinase that exists in two distinct complexes, mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). mTORC1 is composed of mTOR, regulatory associated protein of mTOR (Raptor), mammalian lethal with SEC13 protein 8 (MLST8), PRAS40, and DEPTOR (Kim et al., 2002, Cell 110:163-75; Fang et al., 2001, Science 294(5548):1942-5). mTORC1 integrates four major signaling inputs: nutrients (e.g., amino acids and phosphatidic acid), growth factors (insulin), energy, and stress (e.g., hypoxia and DNA damage). Amino acid availability signals to mTORC1 through a pathway involving Rag and Ragulator (LAMTOR1-3), while growth factors and hormones (e.g., insulin) signal to mTORC1 via Akt, which inactivates TSC2 and prevents mTORC1 inhibition. Alternatively, low ATP levels lead to AMPK-dependent activation of TSC2 and phosphorylation of raptor, reducing mTORC1 signaling proteins.

[0183] Active mTORC1 mediates several metabolic processes, including transcriptional activation leading to mRNA translation via phosphorylation of downstream targets (4E-BP1 and p70 S6 kinase), suppression of autophagy (Atg13, ULK1), ribosome biogenesis, and mitochondrial metabolism or adipogenesis. mTORC1 has several downstream biological effects: mTORC1 activity thus promotes cell proliferation when conditions are favorable, or promotes catabolic processes during stress or when conditions are unfavorable.

[0184] mTORC2 is composed of mTOR, rapamycin-insensitive companion of mTOR (RICTOR), GβL, and mammalian stress-activated protein kinase-interacting protein 1 (mSIN1). In contrast to mTORC1, for which many upstream signals and cellular functions have been defined (see above), relatively little is known about the biology of mTORC2. mTORC2 regulates cytoskeletal organization through its stimulation of F-actin stress fibers, paxillin, RhoA, Rac1, Cdc42, and protein kinase Cα (PKCα). Knockdown of mTORC2 components has been observed to affect actin polymerization and disrupt cell morphology (Jacinto et al. (2004). Nat. Cell Biol. 6:1122–1128; Sarbassov et al. (2004). Curr. Biol. 14:1296–1302). This suggests that mTORC2 regulates the actin cytoskeleton by promoting the phosphorylation of protein kinase Cα (PKCα), the phosphorylation and relocalization of paxillin to focal adhesions, and the GTP loading of RhoA and Rac1. The molecular mechanisms by which mTORC2 regulates these processes remain to be determined.

[0185] In some embodiments, the mTOR inhibitor is an inhibitor of mTORC 1. In some embodiments, the mTOR inhibitor is an inhibitor of mTORC2.

[0186] In some embodiments, the mTOR inhibitor is a limus drug, including sirolimus and its analogs. Examples of limus drugs include, but are not limited to, temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the limus drug is selected from the group consisting of temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506).

[0187] In some embodiments, the mTOR inhibitor is sirolimus, a macrolide antibiotic that inhibits the mTOR pathway by complexing with FKBP-12 and binding mTORC1.

[0188] In some embodiments, the mTOR inhibitor is sirolimus (rapamycin), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), PI-103, Ku-0063794, INK 128, AZD2014, NVP-BGT226, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354 and eforolimus (also known as ridaforolimus or deforolimus).

[0189] BEZ235 (NVP-BEZ235) is an imidazoline-binding protein (mTORC1) catalytic inhibitor. It is a quinoline (imidazoquilonine) derivative (Roper J et al., PLoS One, 2011, 6(9), e25132). Everolimus is a 40-O-(2-hydroxyethyl) derivative of rapamycin that binds the cyclophilin FKBP-12, which also complexes with mTORC1. AZD8055 is a small molecule that inhibits the phosphorylation of mTORC1 (p70S6K and 4E-BP1). Temsirolimus is a small molecule that forms a complex with FK506-binding protein and inhibits mTOR activation when it is in the mTORC1 complex. PI-103 is a small molecule that inhibits activation of the rapamycin-sensitive (mTORC1) complex (Knight et al. (2006) Cell. 125:733-47). KU-0063794 is a small molecule that inhibits mTORC1 phosphorylation at Ser2448 in a dose- and time-dependent manner. INK 128, AZD2014, NVP-BGT226, CH5132799, and WYE-687 are each small molecule inhibitors of mTORC1. PF-04691502 inhibits mTORC1 activity. GDC-0980 is an orally bioavailable small molecule that inhibits class I PI3 kinase and TORC1. Torin 1 is a potent small molecule inhibitor of mTOR. WAY-600 is a potent, ATP-competitive, selective inhibitor of mTOR. WYE-125132 is an ATP-competitive small molecule inhibitor of mTORC1. GSK2126458 is an inhibitor of mTORC1. PKI-587 is a highly potent dual inhibitor of PI3Kα, mTORαγ, and mTOR. PP-121 is a multi-target inhibitor of PDGFR, Hck, mTOR, VEGFR2, Src, and Abl. OSI-027 is a selective and potent dual inhibitor of mTORC1 and mTORC2 with IC50s of 22 nM and 65 nM, respectively. Palomid 529 is a small molecule inhibitor of mTORC1 that lacks affinity for ABCB1 / ABCG2 and has good brain penetration (Lin et al. (2013) Int J Cancer DOI: 10.1002 / ijc.28126 (Published electronically ahead of print)). PP242 is a selective mTOR inhibitor.XL765 is a dual mTOR / PI3k inhibitor of mTOR, p110α, p110β, p110γ, and p110δ. GSK1059615 is a novel dual inhibitor of PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ, and mTOR. WYE-354 inhibits mTORC1 in HEK293 cells (0.2μM-5μM) and HUVEC cells (10nM-1μM). WYE-354 is a potent and specific ATP-competitive inhibitor of mTOR. Deforolimus (ridaforolimus, AP23573, MK-8669) is a selective mTOR inhibitor.

[0190] In some embodiments, the mTOR kinase inhibitor is selected from the group consisting of CC-115 and CC-223.

[0191] Other Components in Nanoparticle Compositions The nanoparticles described herein can be present in a composition containing other drugs, excipients, or stabilizers. For example, certain negatively charged components can be added to increase the stability of nanoparticles by increasing their negative zeta potential. Such negatively charged components include, but are not limited to, bile salts from bile acids such as glycocholic acid, cholic acid, chenodeoxycholic acid, taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid, and dehydrocholic acid; phospholipids, including lecithin (egg yolk)-based phospholipids, include the following phosphatidylcholines: palmitoyloleoylphosphatidylcholine, palmitoyllinoleoylphosphatidylcholine, stearoyllinoleoylphosphatidylcholine, stearoyloleoylphosphatidylcholine, stearoylarachidoylphosphatidylcholine, and dipalmitoylphosphatidylcholine. Other phospholipids include L-α-dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), and other related compounds. Negatively charged surfactants or emulsifiers, such as sodium cholesteryl sulfate, are also suitable as additives.

[0192] In some embodiments, the compositions are suitable for administration to humans. In some embodiments, the compositions are suitable for administration to mammals, such as domestic pets and agricultural animals, in veterinary settings. A wide variety of suitable formulations of nanoparticle compositions exist (see, for example, U.S. Patent Nos. 5,916,596 and 6,096,331). The following formulations and methods are merely exemplary and are in no way limiting. Formulations suitable for oral administration may consist of (a) a liquid solution, such as an effective amount of the compound dissolved in a diluent such as water, saline, or orange juice; (b) capsules, sachets, or tablets, each containing a predetermined amount of the active ingredient as a solid or granules; (c) a suspension in a suitable liquid; and (d) a suitable emulsion. Tablet forms may include one or more of lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, gum acacia, gelatin, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, stearic acid, and other excipients, coloring agents, diluents, buffers, moisturizing agents, preservatives, flavoring agents, and pharmacologically compatible excipients. Lozenge forms may include pastilles containing the active ingredient in a flavoring (usually sucrose and gum acacia or gum tragacanth), and the active ingredient in an inert base such as gelatin and glycerin, or sucrose and gum acacia, emulsions, gels, and the like containing the active ingredient plus excipients such as excipients known in the art.

[0193] Examples of suitable carriers, excipients, and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl- and propylhydroxybenzoates, talc, magnesium stearate, and mineral oil. The formulations may further include lubricating agents, wetting agents, emulsifying and suspending agents, preservatives, sweetening agents, or flavoring agents.

[0194] Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation compatible with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, which may include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The formulations may be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and may be stored under freeze-dried (lyophilized) conditions, requiring only the addition of a sterile liquid vehicle for injection, e.g., water, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the types previously described. Injectable formulations are preferred.

[0195] In some embodiments, the composition is formulated to have a pH range of about 4.5 to about 9.0, including, for example, any of the following pH ranges: about 5.0 to about 8.0, about 6.5 to about 7.5, and about 6.5 to about 7.0. In some embodiments, the composition is formulated to have a pH of about 6 or greater, including, for example, any of the following pH ranges: about 6.5, 7, or 8 (e.g., about 8). The composition can also be made isotonic with blood by adding an appropriate tonicity modifier, such as glycerol.

[0196] Kits, medicaments, compositions, and unit dosage forms The present invention also provides kits, medicaments, compositions and unit dosage forms for use in any of the methods described herein.

[0197] The kits of the invention include one or more containers containing a limus drug-containing nanoparticle composition (or unit dosage form and / or article of manufacture), and, in some embodiments, further include instructions for use according to any of the methods described herein. The kits may also further include instructions for selecting individuals suitable for treatment. The instructions provided with the kits of the invention are typically written instructions on a label or package insert (e.g., a paper sheet included within the kit), although machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.

[0198] For example, in some embodiments, the kit includes a) a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin (e.g., human serum albumin), and b) instructions for administering the nanoparticle composition for the treatment of PEComa. In some embodiments, the kit includes a) a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin (e.g., human serum albumin), and b) instructions for administering (e.g., subcutaneously or intravenously) the nanoparticle composition and other agents for the treatment of PEComa. The nanoparticles and other agents may be present in separate containers or in a single container. For example, the kit may include one separate composition, or two or more compositions, one composition comprising nanoparticles and one composition comprising another agent.

[0199] The kits of the present invention are present 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 buffers and interpretive information. Thus, the present application also provides articles of manufacture that include vials (e.g., sealed vials), bottles, jars, flexible packaging, and the like.

[0200] Instructions for use of the nanoparticle composition generally include information about the dosage, administration schedule, and administration route for the intended treatment. The container may be a unit dose, a bulk package (e.g., a multi-dose package), or a sub-unit dose. For example, a kit may be provided containing a sufficient dose of the mTOR inhibitor (e.g., a limus drug, such as sirolimus) and / or gemcitabine disclosed herein to provide effective treatment for an individual over an extended period, such as 1 week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or longer. The kit may also include multiple unit doses of the mTOR inhibitor (e.g., a limus drug) and pharmaceutical composition and instructions for use, packaged in a quantity sufficient for storage and use in a pharmacy, such as a hospital pharmacy or a compounding pharmacy.

[0201] Also provided are medicaments, compositions, and unit dosage forms useful for the methods described herein. In some embodiments, a medicament (or composition) for use in treating PEComa is provided, comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus drug) and albumin (e.g., human serum albumin).

[0202] Those skilled in the art will recognize that multiple embodiments are possible within the scope and spirit of the present invention. The present invention will now be described in more detail by reference to the following non-limiting examples. The following examples further illustrate the present invention but, of course, should not be construed as in any way limiting its scope. [Example]

[0203] Example 1 A Phase II multicenter study with patients receiving ABI-009 treatment Patients with advanced malignant PEComa who had not been previously treated with an mTOR inhibitor were enrolled in a Phase II, single-arm, open-label, multicenter study to evaluate the efficacy and safety profile of intravenous ABI-009 (also referred to herein as Nab-sirolimus or Nab-rapamycin). Stage 2 will accept enrollment if futility criteria are not met.

[0204] At least 30 patients will be enrolled in the study. Malignant PEComa histology may be assessed locally at each institution for enrollment, but all patients must be retrospectively confirmed by centralized review after enrollment to meet pre-specified criteria for malignant PEComa, as outlined in the inclusion criteria. If a patient does not retrospectively meet these criteria, replacement will be considered.

[0205] Patients were eligible for inclusion in this study only if all of the following criteria were met: (i) patients must have a histologically confirmed diagnosis of progressive malignant PEComa; (ii) patients must have available tumor blocks and / or fresh biopsies with corresponding pathology reports (or approximately 30 unstained slides, with a minimum of 16 slides mandatory) to allow for retrospective centralized confirmation of malignant PEComa and mTOR pathway and biomarker analysis; (iii) patients must have one or more target lesions measurable by CT scan or MRI, or disease measurable by RECIST v1.1; (iv) patients must not have been previously treated with an mTOR inhibitor; (v) prior treatment (investigational or otherwise), chemotherapy, radiation therapy, surgery, or other therapeutic agents (other than mTOR inhibitors) are allowed after completion of 5 half-lives or ≥ 28 days, whichever is shorter, prior to enrollment; (vi) eligible patients, aged 18 years or older, must be enrolled in a clinical trial with an MRSA-associated or non-mTOR inhibitor ... (vii) Patients must have the following blood chemistry levels at screening (obtained (local laboratory) ≤14 days prior to enrollment): (a) total bilirubin ≤1.5 × upper limit of normal (ULN) mg / dl, (b) AST ≤2.5 × ULN (≤5 × ULN if due to liver metastases), (c) serum creatinine ≤1.5 × ULN; (viii) adequate biological parameters at screening as demonstrated by the following blood counts (obtained ≤14 days prior to enrollment, local laboratory): (a) absolute neutrophil count (ANC) ≥1.(b) platelet count ≥ 100,000 / mm3 (100 × 109 / L), (c) hemoglobin ≥ 9 g / dL; (ix) urine protein < 2 g proteinuria / 24 hours; (x) serum triglycerides < 300 mg / dL; serum cholesterol < 350 mg / dL; (xi) males or non-pregnant, non-lactating females (females of childbearing potential must agree to use effective contraception without interruption for 28 days prior to starting IP during study medication, have a negative serum pregnancy test (β-hCG) result at screening, and maintain a normal pregnancy status throughout the course of the study and beyond). (xii) a life expectancy of >3 months as determined by the investigator; (xiii) the ability to understand and sign informed consent; (xiv) willingness and ability to comply with scheduled clinic visits, laboratory tests, and other study procedures.

[0206] Patients are not eligible for inclusion in this study if any of the following criteria apply: (i) patients with lymphangioleiomyomatosis (LAM) are excluded; (ii) known active, uncontrolled or symptomatic central nervous system (CNS) metastases (patients with controlled and asymptomatic CNS metastases may participate in this study, provided that they have completed any prior treatment for CNS metastases (including radiation therapy and / or surgery) ≥28 days prior to the start of treatment in this study and should not be receiving chronic corticosteroid treatment for CNS metastases); (iii) active gastrointestinal bleeding if infusion-dependent; (iv) pre-existing thyroid abnormalities are permitted, provided that thyroid function can be controlled with medication; (v) uncontrolled serious medical or psychiatric illness.Patients with "currently active" second malignancies other than non-melanoma skin cancer, cervical carcinoma in situ, resected incidental prostate cancer (staged pT2 with a Gleason score ≤6 and a postoperative PSA <0.5 ng / mL) or other adequately treated carcinoma in situ are ineligible (patients are not considered to have "currently active" malignancies if they have completed treatment and have been disease-free for ≥1 year); (vi) liver-directed treatments within 2 months of enrollment (including prior treatment with radiation therapy (including radiolabeled spheres and / or CyberKnife), hepatic artery embolization (with or without chemotherapy), or cryotherapy / ablation) are acceptable if these treatments did not affect the area of ​​measurable disease used in this protocol. (vii) recent infection requiring systemic anti-infective treatment completed ≤14 days prior to enrollment (excluding uncomplicated urinary tract infection or upper respiratory tract infection); (viii) uncontrolled diabetes mellitus defined by HbA1c>8% despite appropriate treatment, unstable coronary artery disease or myocardial infarction within the preceding 6 months; (ix) receiving any concomitant anti-tumor treatment; (x) patients with a history of interstitial lung disease and / or interstitial pneumonia, or pulmonary hypertension; (xi) the use of certain medications and illicit drugs within 5 half-lives or 28 days, whichever is shorter, prior to the first dose of study drug and for the duration of the study is not permitted; (xii) the use of strong inhibitors and inducers of CYP3A4 within 14 days prior to receiving the first dose of ABI-009. Additionally, use of any known CYP3A4 substrate with a narrow therapeutic window (e.g., fentanyl, alfentanil, astemizole, cisapride, dihydroergotamine, pimozide, quinidine, terfanide) within 14 days prior to receiving the first dose of ABI-009.

[0207] The study is expected to last approximately 32 months from first patient enrollment to last patient follow-up, including an approximately 24-month enrollment period, an estimated 6 months of treatment (or until treatment is no longer tolerated), and an end of treatment visit 4 weeks (+ / - 7 days) after the last treatment.

[0208] The following assessments will be performed on Day 1 of each cycle unless otherwise specified: (i) physical examination; (ii) weight assessment; (iii) BSA calculation; (iv) evaluation of concomitant medications and procedures; (v) vital signs (e.g., temperature, systolic and diastolic blood pressure, and pulse); (vi) ECOG performance status; (vii) ECG; (viii) clinical chemistry panel (including, but not limited to, sodium, potassium, chloride, glucose, alkaline phosphatase (ALP), AST / SGOT, ALT / SGPT, serum albumin); (ix) CBC, differential, and platelet count; (x) thyroid function; (xi) screening for hepatitis and HIV infection (all odd-numbered cycles starting on C3); (xii) fasting lipids (all even-numbered cycles starting on C2); (xiii) adverse event assessment; and (xiv) pharmacokinetic assessment (Day 1 of Cycle 1 only).

[0209] The following assessments will be performed on Day 8 of each cycle unless otherwise specified: (i) assessment of concomitant medications and procedures; (ii) vital signs; (iii) CBC, differential, and platelet count; (iv) thyroid function; (v) adverse event assessment; (vi) pharmacokinetic assessment (Day 8 of Cycle 1). only).

[0210] Tumor response will be assessed by CT or MRI scans of the chest, abdomen, and pelvis; image preparation and evaluation will follow the specifications provided in RECIST version 1.1. The same modality (CT or MRI) must be used at screening and throughout the study.

[0211] End of study (EOS) is defined as either the date of the last patient's last visit to complete the study or the date of receipt of the last data point from the last patient required for the primary, secondary, and / or exploratory analyses, as pre-specified in the protocol.

[0212] The end of treatment (EOT) for a patient is defined as the date of the last dose of ABI-009. The end of treatment visit for a patient is when safety assessments and procedures are performed after the last treatment, which must occur at least 4 weeks (± 7 days) after the last dose of ABI-009.

[0213] The follow-up period is the on-study period after the EOT visit. All patients who discontinue study drug and have not withdrawn full consent to participate in the study will continue the follow-up phase for survival and initiation anti-cancer treatment. Follow-up will continue approximately every 12 weeks (+ / - 3 weeks) until death, withdrawal of consent, or end of study, whichever occurs first. This assessment may be performed by record review and / or telephone contact.

[0214] A rapamycin PK study will be conducted with limited PK sampling for all patients in this Phase 2 study. Blood samples will be obtained only during Cycle 1 Day 1 (C1D1) and will be collected immediately prior to the dose (pre-infusion), 0.5 hours (immediately prior to the end of the infusion), 1 hour, 2 hours, 4 hours, and 168 hours (immediately prior to the C1D8 dose). Note that T=0 is defined as the start of the infusion; i.e., all sample collection times are relative to the start of the infusion. The end-of-infusion (0.5 hour) sample will be collected immediately prior to stopping the infusion. If the duration of the infusion is varied, samples should be collected immediately prior to the end of the infusion. Whole blood samples will be collected into EDTA tubes for rapamycin determination in a central laboratory.

[0215] Patients will receive ABI-009 at 100 mg / m² via 30-minute IV infusion for 2 weeks every 3 weeks. Two dose reduction levels will be allowed: 75 mg / m² and 56 mg / m². Patients will continue treatment until disease progression, unacceptable toxicity, until in the investigator's opinion the patient is no longer benefiting from treatment, or at the patient's discretion.

[0216] Patients will be evaluated for complete response (CR), partial response (PR), stable disease (SD), or progressive disease (PD) by CT imaging. Contrast-enhanced MRI may also be used as long as the same modality is used throughout the study. Baseline scan results may be received from an outside facility but must be performed within 4 weeks of the start of treatment and must include a CT or MRI of the chest, abdomen, and pelvis (if clinically indicated). The first response assessment, with a CT or MRI scan reporting target lesions, should be performed 6 weeks after the first treatment and repeated every 6 weeks for the first year, then every 12 weeks thereafter until disease progression. If an initial observation of objective response (CR or PR) is made, a confirmatory scan should be performed 6 weeks after the initial observation.

[0217] The primary endpoint, ORR, will be determined by independent radiologist(s). Independent radiological review will follow a separate imaging protocol.

[0218] After disease progression, patients will be followed for survival every 12 weeks or more frequently as needed until death, withdrawal of consent, or end of study, whichever occurs first.

[0219] Safety and tolerability will be monitored through ongoing reporting of treatment and treatment-related adverse events (AEs), particularly AEs of interest, laboratory abnormalities, and the incidence of patients experiencing dose modifications, dose delays / missed doses, dose interruptions, and / or premature discontinuation of IP due to AEs. All AEs will be recorded by the investigator from the time the patient signs the informed consent until 28 days after the last dose of IP. Adverse events will be graded according to the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) v4.03.

[0220] Physical examination, vital signs, laboratory assessments (e.g., serum chemistry, hematology), and ECOG performance status will be monitored. All SAEs (whether related to IP or not) will be followed until resolution. Laboratory analyses will be performed according to the study schedule.

[0221] The primary analysis will be performed when all study subjects have been treated and all patients have had an opportunity to be treated for at least 6 months. All primary, secondary, and exploratory efficacy and safety analyses will be performed at the time of the primary analysis, except for biomarkers that may be analyzed at a later date.

[0222] The primary endpoint was ORR by independent radiological review, defined as the proportion of patients achieving a confirmed PR or CR according to RECIST 1.1.

[0223] This study is designed to test whether the confirmed ORR in patients treated with ABI-009 is greater than 5%, as determined using a one-sided exact binomial test with a type I error of 0.025. With approximately 30 patients, this study provides greater than 95% power to reject the null hypothesis that the ORR is ≦5%, assuming a true response rate of 30%. In addition to hypothesis testing, the number and percentage of patients achieving the ORR will be summarized, providing an exact 95% confidence. Assuming an observed ORR of 30%, the lower bound of the 95% confidence interval (CI) for the estimated ORR excludes values ​​less than 14.7%.

[0224] Analyses of the secondary efficacy endpoints of PFS, 6-month PFS rate, DOR, and OS will be performed separately for two subgroups of patients: 1) patients with metastatic disease; and 2) patients with unresectable locally advanced disease or resectable disease with multiple resections. Some patients in the locally advanced subgroup may be clinically indicated to undergo surgery if there is sufficient tumor shrinkage, which introduces bias into the analysis.

[0225] For patients with metastatic disease, PFS at 6 months, median PFS, DOR, and OS are summarized using Kaplan-Meier (KM) analysis. Quartiles are summarized with 95% CI. The number of patients with unresectable locally advanced disease or resectable disease with multiple resections is expected to be small; therefore, PFS (median and 6 months), DOR, and OS for these patients are summarized by descriptive statistics.

[0226] An important goal of tumor molecular profiling is to identify specific markers that predict response to ABI-009.

[0227] Blood samples were collected for cell-free plasma DNA collection (pre- and post-treatment samples, mandatory): Cell-free plasma DNA assays are obtained for molecular analysis using next-generation sequencing to assess the prevalence of identified mutations over time in primary tumor samples as a measure of response.

[0228] Tumor biopsy mutation analysis or epigenetic changes will also be completed. Archival or fresh tissue biopsies from pre-treatment tumors will be required (mandatory) for patients in this study. Additionally, if available, intra-treatment biopsies will be performed to assess pharmacodynamic effects. Tumor samples will be collected after the last treatment and at the time of progression, if biopsies are performed. Tumor biopsies will be collected and analyzed for pharmacodynamic changes to determine the drug's effect on target(s) in the tumor and potentially analyze acquired resistance and associated molecular mechanisms. Exome sequencing of approximately 300 genes using the ONCOPANEL™ test (BWH Pathology Department, CLIA-certified) will be performed to evaluate mutations in all known mTOR pathway genes, including, but not limited to, PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR, and RHEB. Evaluation of correlation of clinical response to treatment will be performed, testing the correlation between biopsy and circulating DNA analysis. FISH (fluorescence in situ hybridization) analysis of translocations in TFE3 will be completed. These studies will be conducted at the Center of Advanced Molecular Diagnostics laboratory at Brigham and Women's Hospital, Boston, Massachusetts, which is also CLIA-certified.

[0229] Immunohistochemistry is completed for relevant pathway markers, including but not limited to phosphoproteins p-AKT, p-S6, p-S6K, p-4EBP1, and p-SPARC; proliferation markers, such as Ki-67; and apoptosis markers, such as PARP or its fragments. Post-treatment (progression) samples are analyzed by exome sequencing similar to that described above to explore causes of resistance, including secondary mutations and genomic amplification or deletion events.

[0230] Example 2 A clinical pilot study of Nab-sirolimus in mTOR pathway-altered malignancies A single-arm, Phase II clinical trial is designed to evaluate the efficacy of Nab-sirolimus (also referred to as ABI-009) in individuals with PEComa and mTOR pathway gene mutation status, particularly those with gene mutation status that confers sensitivity to mTOR inhibitors (e.g., a composition comprising nanoparticles containing a limus drug and albumin). Gene mutation status will be identified through next-generation sequencing experiments from individuals in the clinical study. The primary objective of the study is to evaluate the response rate of Nab-sirolimus in individuals with PEComa and mTOR-activating gene mutation status. Secondary objectives are (1) to estimate the time to progression and overall survival of selected individuals; and (2) to estimate the adverse event profile of Nab-sirolimus in selected individuals. Additionally, a correlational study will be conducted to assess the proportion of individual mTOR-activating gene mutation status and to evaluate the association between individual mTOR-activating gene mutation status and clinical outcomes for individuals with PEComa.

[0231] A single group of individuals will be enrolled in the clinical study. Prior to enrollment, individuals will be evaluated in a CLIA-certified laboratory for mTOR-activating gene mutations in at least one mTOR-related gene selected from the group consisting of AKT1, MTOR, PIK3CA, PIK3CG, TSC1, TSC2, RHEB, STK11, NF1, NF2, PTEN, TP53, FGFR4, and BAP1. Individuals with at least one mTOR-activating gene mutation and meeting all inclusion criteria will be selected for treatment. Archival paraffin-embedded (PPFE) tissue samples may optionally be obtained from each individual. Selected individuals will receive intravenous Nab-sirolimus at a dose of approximately 75 mg / m² on days 1, 8, and 15 of a 28-day cycle, or approximately 100 mg / m² on days 1 and 8 of a 21-day cycle. Nab-sirolimus will be infused over approximately 30 minutes between each administration. Individuals continue to receive Nab-sirolimus treatment and are actively monitored until disease progression and / or the occurrence of intolerable adverse events occur, or until the individual refuses to receive treatment.If multiple adverse events are observed, the dose of Nab-sirolimus may be interrupted or reduced to allow drug-related toxicity management.For example, the dose of Nab-sirolimus may be first reduced to 56 mg / m2 IV on days 1, 8, and 15 of a 28-day cycle, and then reduced a second time to 45 mg / m2 IV on days 1, 8, and 15 of a 28-day cycle.Only two dose reductions are permitted per individual.Supportive treatments, such as antiemetics, growth factors (G-CSF), bisphosphonates or denosumab for existing painful bone metastases, blood and blood products, warfarin or LMWH, and / or loperamide for diarrhea, may be permitted at the physician's discretion. These individuals should be returned to the consenting facility for treatment and evaluated at least every 28 days (or about every 25 to about 31 days) during treatment.Biomarkers (e.g., sequences and levels of AKT1, MTOR, PIK3CA, TSC1, TSC2, RHEB, STK11, NF1, NF2, and PTEN, and levels of phosphorylated 4EBP1 and S6) will be assessed for each individual on day 1 of cycle 1, day 1 (±3 days) of cycle 2, and day 1 (±3 days) of cycle 3, and then every two cycles thereafter. Blood samples will be collected from each individual to analyze circulating (e.g., cell-free) DNA before and after the entire course of treatment.

[0232] Various biological samples are collected from each individual during the course of the study (e.g., before, during, and after treatment), and these biological samples are used to assess the mutation status and levels of relevant biomarkers. In-treatment biological samples may be collected from individuals, for example, on day 1 of cycle 1, day 1 (±3 days) of cycle 2, and day 1 (±3 days) of cycle 3, and then every two cycles thereafter. Blood samples are collected from each individual before and after treatment. Cell-free plasma DNA samples are prepared from each blood sample for evaluation of circulating DNA. The cell-free plasma DNA samples are analyzed using next-generation sequencing to assess the prevalence of mTOR-activating gene mutations identified in primary tumor samples over time as a measure of response to treatment. In addition, fresh or archived (e.g., PPFE) tumor biopsy samples are collected from each individual before treatment and, optionally, during the course of treatment (i.e., during treatment). In-treatment tumor biopsy samples are used to assess the pharmacodynamic effects of Nab-sirolimus in individuals. After treatment, tumor biopsy samples are collected from each individual at the time of disease progression after response to treatment to evaluate the mechanism of resistance, including secondary mutation, genome amplification or gene deletion events.Exome sequencing experiments are carried out using ONCOPANEL™ test for approximately 300 genes to evaluate the mutations in mTOR pathway genes, including but not limited to PIK3CA, TSC1, TSC2, AKT, PTEN, MTOR and RHEB.In addition, mTOR activation abnormalities (for example, the sequence and level of biomarkers, including but not limited to AKT1, MTOR, PIK3CA, PIK3CG, TSC1, TSC2, RHEB, STK11, NF1, NF2, PTEN, TP53, FGFR4 and BAP1), and the levels of phosphorylated AKT (i.e., p-AKT), 4EBP1 (i.e., p-4EBP1) and S6K (i.e., p-S6K)) are evaluated using tumor biopsy samples. Proliferative markers (e.g., Ki-67) and apoptotic markers (e.g., PARP) may be assessed using immunohistochemistry. FISH (fluorescence in situ hybridization) analysis of translocations in TFE3 is performed.The results of the evaluation will be used to assess the correlation between mTOR-activating gene mutations and clinical response to treatment, and to test the correlation between mTOR-activating gene mutations identified in tumor biopsies and circulating DNA.

[0233] The primary endpoint of this study is the rate of confirmed response. In PEComa, a confirmed response is defined as either a complete response or partial response, noted as an objective outcome on two consecutive assessments at least 8 weeks apart. Confirmed response will be assessed during every cycle of treatment. Exact binomial confidence intervals for the true confirmed response rate will be calculated. Secondary endpoints of this study include survival time, time to disease progression, and adverse events. The distributions of survival time and time to disease progression will be estimated using the Kaplan-Meier method. For all primary and secondary endpoints, statistical analyses will be performed for the entire patient population and within each disease group.

[0234] A correlation study will be conducted to determine the association between treatment and quality of life and individual mTOR-activating gene mutation status for the entire patient group. Quality of life will be assessed before examining treatment response since the last treatment evaluation and considering the patient's overall health. Quality of life will be measured using the EORTCQLQ-C30, a 30-item patient-reported questionnaire, regarding the patient's ability to function, symptoms related to cancer and its treatment, overall health and quality of life, and the perceived financial impact of cancer and its treatment. Scale score trajectories of quality of life over time will be examined using stream plots and mean plots with standard deviation error bars. Changes from baseline at each cycle are statistically tested using paired t-tests, and standardized response means are interpreted after applying Middle (2002) adjustments using Cohen's (1988) cutoffs: <0.20 = mediocre; 0.20-<0.50 = small; 0.5-<0.8 = moderate; and ≥0.8 = large. Rates of individual mTOR activation abnormalities are described, and associations with confirmed response are examined using Fisher's exact test. Associations with time to progression and overall survival are examined using the log-rank test. A one-sided p-value ≤0.10 is considered statistically significant from start to finish.

[0235] Eligible individuals must meet all of the following inclusion criteria: (a) have histological confirmation of PEComa (e.g., lymphangioleiomyomatosis); (b) have advanced stage cancer; (c) have at least one mTOR pathway gene mutation confirmed in a CLIA-certified laboratory, where the mTOR pathway mutation is one of the following: genetic mutations in AKT1, MTOR, PIK3CA, TSC1, TSC2, RHEB, STK11, NF1, NF2, PTEN (e.g., PTEN deletion), TP53, FGFR4, and BAP1. (d) not having received any of the following treatments: (1) chemotherapy within 4 weeks prior to treatment with Nab-sirolimus; (2) hormonal therapy within 4 weeks prior to treatment with Nab-sirolimus; (3) radiation therapy within 4 weeks prior to treatment with Nab-sirolimus; (4) treatment with nitrosoureas, mitomycin, or extensive radiation therapy within 6 weeks prior to treatment with Nab-sirolimus; (5) immunosuppressants (excluding corticosteroids used as antiemetics) within 3 weeks prior to treatment with Nab-sirolimus. (6) use of prior mTOR pathway inhibitor therapy; (d) having the following laboratory values ​​obtained 14 days or less prior to enrollment: (1) absolute neutrophil count (ANC) ≥ 1500 / mm3, platelet count ≥ 100,000 / mm3; (2) hemoglobin ≥ 9.0 g / dL; (3) total bilirubin ≤ 1.5 × institutional upper limit of normal (ULN); (4) aspartate transaminase (AST); alanine aminotransferase (ALT) ≤ 3 × ULN, or ≤ 5 × ULN if the subject has tumor lesions in the liver; (5) serum cholesterol ≤ (6) serum triglycerides ≤ 300 mg / dL; (7) serum creatinine ≤ 1.5 × ULN; (e) have previously failed, tolerated, or refused other available active treatments; (f) have adequate coagulation function as defined by any of the following criteria: (1) INR ≤ 1.5 × ULN; (2) for subjects receiving warfarin or LMWH, these subjects must, in the opinion of the investigator, be clinically stable without evidence of active bleeding while on anticoagulant therapy.

[0236] Exclusion criteria were: (a) pregnant or nursing women, or women of childbearing potential who are biologically capable of becoming pregnant, or men capable of fathering children who are not using two forms of highly effective contraception; (b) patients with a history of interstitial lung disease and / or pneumonia; (c) receiving any concomitant antitumor treatment or inhibitor of CYP3A4; (d) antibiotics with similar chemical or biological composition, including macrolide antibiotics (e.g., azithromycin, clarithromycin, dirithromycin, and erythromycin) and ketolide antibiotics. (e) major surgery (e.g., intrathoracic, intraperitoneal, or intrapelvic) ≤ 4 weeks prior to enrollment, or inability to recover from the side effects of such surgery, excluding port placement, nephrectomy, tumor biopsy, and minor surgery; (f) concomitant use of any other approved or investigational anticancer agent considered for treatment of a primary neoplasm; (g) uncontrolled diabetes mellitus as defined by HbA1c > 8% despite appropriate therapy; (h) unstable coronary artery disease or myocardial infarction during the preceding 6 months; and (i) hypertension uncontrolled by medical therapy.

[0237] Example 3 Treating Metastatic PEComa with nab-sirolimus A patient with pulmonary PEComa (pretreated with additional radiation therapy and then surgically removed approximately 5 months earlier) presented with a new, suspicious metastatic lesion on a CT scan. This new precarinal (mediastinal) lymph node measured approximately 1.5 cm in diameter. Because the size of the new lesion was borderline for the entry criteria for PEC001 (RECIST, requiring measurable disease), the patient underwent another CT scan approximately 1 month later. At that time, the lesion was measurable, and the patient also had a new pleural lesion measuring >2 cm. Given the presence of a new lesion since the last CT scan, the metastatic PEComa was considered a rapidly progressing, invasive tumor. The patient consented to the PEC001 study and treatment with nab-sirolimus (also referred to as ABI-009), starting at a dose of 100 mg / m2 given by IV infusion over 30 minutes, repeated on a 2-weekly schedule every 3 weeks. The next follow-up CT scan was scheduled for 6 weeks later according to the PEC001 clinical protocol. During that period, the patient received nab-sirolimus according to the protocol and tolerated the drug well, with no significant adverse events reported. At the 6-week follow-up CT scan, these lesions showed no change from the previous scan, and the disease was considered to have been kept stable by treatment with nab-sirolimus. Further treatment and follow-up continued according to the protocol, and the patient continued to tolerate the treatment without significant adverse events.

Claims

1. A composition comprising nanoparticles comprising sirolimus and albumin for treating perivascular epithelioid cell tumor (PEComa) in an individual, wherein the individual is selected for treatment based on their TFE3 and / or TP53 mutation status.

2. 2. The composition of claim 1, wherein the PEComa is pulmonary clear cell sugar tumor, PEComa not otherwise specified (PEComa-NOS), angiomyolipoma, or lymphangioleiomyomatosis.

3. The composition of claim 1 or 2, wherein the PEComa is malignant.

4. 4. The composition of any one of claims 1 to 3, wherein the PEComa is progressive, locally advanced, metastatic, recurrent, or locally unresectable.

5. 2. The composition of claim 1, wherein the PEComa is located in or closely associated with the uterus, kidney, bladder, or prostate.

6. The composition of claim 5 , wherein the PEComa is located within or closely associated with the uterus.

7. The composition of claim 1 , wherein the individual is at least 50 years old.

8. The composition of claim 1 , wherein the composition is administered in a neoadjuvant setting.

9. The composition of claim 1 , which is administered intravenously.

10. 10. The composition of claim 1, wherein the composition is administered intravenously by infusion over 30 minutes.

11. A composition comprising nanoparticles comprising sirolimus and albumin for treating malignant perivascular epithelioid cell tumor (PEComa) in an individual, wherein the individual is at least about 50 years old.

12. A composition comprising nanoparticles comprising sirolimus and albumin for treating malignant perivascular epithelioid cell tumor (PEComa) in an individual, wherein the composition is administered intravenously and the malignant PEComa is localized and unresectable.

13. A composition comprising nanoparticles comprising sirolimus and albumin for treating malignant perivascular epithelioid cell tumor (PEComa) in an individual, the composition being administered intravenously by infusion over 30 minutes.

14. A composition comprising nanoparticles comprising sirolimus and albumin for treating malignant perivascular epithelioid cell tumor (PEComa) in an individual, wherein the composition is administered intravenously and in a neoadjuvant setting.

15. The effective amount of sirolimus in the nanoparticle composition is about 10 mg / m ２ ~Approx. 150mg / m ２ 15. The composition of any one of claims 1 to 14, wherein

16. The effective amount of sirolimus in the nanoparticle composition is about 100 mg / m ２ 16. The composition of any one of claims 1 to 15, wherein

17. 17. The composition of any one of claims 1 to 16, wherein the nanoparticle composition is administered weekly during treatment.

18. 18. The composition of any one of claims 1 to 17, wherein the nanoparticle composition is administered weekly for two weeks out of every three weeks during treatment.

19. 20. The composition of claim 18, wherein the nanoparticle composition is administered on days 1 and 8 of a 21-day cycle during treatment.

20. 20. The composition of any one of claims 11 to 19, wherein the nanoparticle composition is administered intravenously.

21. The composition of any one of claims 11 to 19, wherein the nanoparticle composition is administered subcutaneously.

22. 22. The composition of claim 1, wherein the nanoparticles in the composition have an average diameter of about 120 nm or less.

23. 23. The composition of any one of claims 1 to 22, wherein the sirolimus in the nanoparticles is associated with the albumin.

24. 24. The composition of any one of claims 1 to 23, wherein the weight ratio of albumin to sirolimus in the nanoparticle composition is from about 1:1 to about 9:

1.

25. 25. The composition of any one of claims 1 to 24, wherein the albumin is human serum albumin.

26. 26. The composition of any one of claims 1 to 25, wherein the individual is a human.

27. 27. The composition of any one of claims 1 to 26, wherein the individual is selected for treatment based on the levels of melanocyte and smooth muscle markers.

28. 28. The composition of claim 27, wherein the melanocyte marker is selected from the group consisting of HMB45, MelanA, and microphthalmia transcription factor.

29. 28. The composition of claim 27, wherein the smooth muscle marker is selected from the group consisting of smooth muscle actin, pan-muscle actin, h-caldesmon, and calponin.

30. 30. The composition of any one of claims 1 to 29, wherein the individual has not been previously treated with an mTOR inhibitor.

31. 31. The composition of any one of claims 1 to 30, wherein the individual has been previously treated with chemotherapy, radiation, or surgery.

32. 30. The composition of any one of claims 1 to 29, wherein the individual has been previously treated with an mTOR inhibitor.

33. 33. The composition of claim 32, wherein the mTOR inhibitor of said treatment is in a non-nanoparticulate formulation.

34. 34. The composition of claim 32 or 33, wherein the individual is resistant or progressive to prior mTOR inhibitor treatment.