Described herein are methods and compositions for treating cancer. Aspects of the invention relate to administering to a subject having cancer an asparaginase and an agent that inhibits GSK3f. Another aspect of the invention relates to administering an asparaginase to a subject having cancer that comprises an inactivating mutation in GSK3f.

The disclosure provides, in various embodiments, polypeptides that upon administration to a mammalian subject, elicit a reduced immunogenicity in the subject, relative to a bacterial asparaginase. The disclosure also provides, in various embodiments, fusion proteins comprising one or more of the polypeptides, polynucleotides encoding the polypeptides, vectors and host cells suitable for expressing the polypeptides, methods for treating a disease (e.g., cancer) in mammalian subject (e.g., a human), and methods for reducing a level of asparagine or asparagine-containing product in a biological fluid from a mammalian subject.

The present invention relates to novel dendrimer conjugates, methods for their preparation and their use for treatment of diseases. The invention discloses a new method for the delivery of dendrimer conjugates with therapeutically active molecules into the cell by utilizing transmembrane solute carrier proteins enabling uptake of the inventive dendrimer conjugates. Particular subject-matter of the present invention is a conjugate of the formula E-[G-L-D(OSO.sub.3.sup.−M.sup.+).sub.n].sub.m, wherein E is a therapeutic or diagnostic effector molecule, wherein D(OSO.sub.3.sup.−M.sup.+).sub.n is a dendrimer D carrying a number n of sulfate groups OSO.sub.3.sup.−M.sup.+, wherein the number n of sulfate groups is selected from 6 to 96, wherein M is a cationic inorganic or organic counter ion to the anionic sulfate group, wherein L is a linker or spacer between D and E, wherein G is a connecting functional group forming the attachment between L and E, and wherein m is an integer from 1 to 20.

Provided herein are methods of production of recombinant E. coli asparaginase. Methods herein allow production of asparaginase in Pseudomonadales host cells at high expression levels and having activity comparable to commercially available asparaginase preparations.

The present disclosure discloses a thermophilic L-asparaginase mutant and screening and fermentation methods thereof, and belongs to the field of gene engineering, enzyme engineering and fermentation engineering. In Bacillus subtilis 168, a Pyrococcus yayanosii CH1-derived L-asparaginase encoding gene is used as a template, and a mutation library is constructed by an error-prone PCR (epPCR) technology. A mutant strain with improved specific enzyme activity is screened through a high-flux screening method of synchronous cell disruption and enzyme activity measurement. Mutated residues included in a positive mutant are analyzed to construct a composite mutant strain S17G/A90S/R156S/K272A with improved specific enzyme activity and specific enzyme activity of 3108 U/mg. An expression quantity of the composite mutant strain in the Bacillus subtilis 168 is increased through measures of a strong promoter P.sub.43 and RBS optimization. Finally, the Bacillus subtilis 168 with a gene of the L-asparaginase composite mutant strain is subjected to enzyme production fermentation in a 5 L fermentation tank through culture medium optimization and pH and feeding coupling strategies. The enzyme activity yield of the L-asparaginase is up to 6453+/−127 U/mL.

A novel protein deamidase having an activity of directly acting on a side chain amide group of an asparagine residue in a protein to form a side chain carboxyl group and release ammonia, a microorganism that produces the same, a gene encoding the same, a method for producing the same, and use of the same are provided. A bacterium classified into the class Actinobacteria is cultured to generate protein deamidase, and the enzyme is collected from culture.

Peptides that specifically bind erythrocytes are described. These are provided as peptidic ligands having sequences that specifically bind, or as antibodies or fragments thereof that provide specific binding, to erythrocytes. The peptides may be prepared as molecular fusions with therapeutic agents, tolerizing antigens, or targeting peptides. Immunotolerance may be created by use of the fusions and choice of an antigen on a substance for which tolerance is desired.

Provided is an in vitro method for determining the metabolic status of a lymphoma comprising a step of determining the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in lymphoma cells, wherein a low level of GAPDH expression is indicative of oxidative phosphorylation (OXPHOS) status. Also provided is an in vitro method for predicting the responsiveness of a patient afflicted with a lymphoma to a treatment with a metabolic inhibitor selected from the group consisting of mitochondrial metabolic inhibitors and glutamine metabolism inhibitors comprising a step of determining the level of GAPDH expression in lymphoma cells obtained from said patient, wherein a low level of GAPDH expression is predictive of a response to a treatment with a metabolic inhibitor.

Provided are various embodiments relating to variant asparaginase polypeptides with enhanced stability, pharmacodynamics, and/or reduced immunogenicity. The variant asparaginase polypeptides may be used as therapeutics in mammals, including human, canines, felines, and equines.

Provided herein are methods of production of recombinant Erwinia asparaginase. Methods herein produce asparaginase having high expression levels in the periplasm or the cytoplasm of the host cell having activity comparable to commercially available asparaginase preparations.