# US 2025/0154601 A1 · Method for hormone or drug screening in a tissue sample > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0154601 A1 - Document ID: US20250154601A1 - Status: Published application - Kind: A1 - Application number: 18/840,661 - Filed: 2023-02-21 - Published: 2025-05-15 - Earliest claimed: 2022-02-22 - Assignees: Ecole Polytechnique Federale de Lausanne - Applicants: Ecole Polytechnique Federale de Lausanne (EPFL) - Inventors: Yueyun ZHANG; Christoph MERTEN; Cathrin BRISKEN - Web page: https://patentgrep.com/patent/US20250154601A1 ## CPC - C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575) - C12N 5/0012: {Cell encapsulation} - C12N 5/0693: {Tumour cells; Cancer cells} - C12N 2503/02: Drug screening - C12N 2513/00: 3D culture - C12N 2533/74: Alginate - C12Q 2600/106: Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism - C12Q 2600/136: Screening for pharmacological compounds - C12Q 2600/158: Expression markers - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to a method for analyzing cellular responsiveness to hormones or drags in a tissue sample based on an alginate-based 3D ex vivo culture system for normal and malignant tissue microstructures and uses thereof. ## Claims ### Claim 1 A method for analyzing cellular responsiveness to hormones or drugs in a tissue sample from a patient or in a patient-derived tissue sample, comprising the steps of: a) Mechanical and/or enzymatic digestion of said tissue sample to produce tissue microstructures; b) Encapsulation of said tissue microstructures in alginate; c) Exposing the encapsulated tissue microstructure to hormones or drugs; d) Cell lysis, mRNA capture and reverse transcription in one-step; e) Alginate dissociation; f) RNA sequencing; wherein step d) is performed with the alginate still crosslinked and intact. ### Claim 2 (depends on 1) The method according to claim 1, wherein in step a) the proteins from the extracellular matrix and cells from stroma of the tissue sample are preserved as a natural surround environment for the microstructures. ### Claim 3 (depends on 1) The method according to claim 1, wherein in step b) no supplementary hormones or growth factors are added during the encapsulation. ### Claim 4 (depends on 1) The method according to claim 1, wherein in step b) no collagen-based biomaterial is added during the encapsulation. ### Claim 5 (depends on 1) The method according to claim 1, wherein in step b) the tissue microstructures are encapsulated in alginate beads having a diameter of between 10 to 1200 μm, preferably between 40 to 800 μm, more preferably between 50 to 500 μm. ### Claim 6 (depends on 1) The method according to claim 1, wherein the encapsulated tissue microstructures are maintained in culture for 3 h to 23 days, preferably 5 h to 14 days, more preferably 12 h to 10 days, even more preferably 24 h to 7 days prior to step c). ### Claim 7 (depends on 1) The method according to claim 1, wherein in step b) encapsulated tissue microstructures are in static culture or cultured without agitation. ### Claim 8 (depends on 1) The method according to claim 1, wherein said tissue sample is selected from the group consisting of epithelium tissue, connective tissue, cartilage tissue, bone tissue, muscle tissue, nerve tissue, blood vessel tissue, heart tissue, lymphatic system tissue, respiratory tract tissue, oesophagus tissue, urinary tract tissue, endocrine system tissue, reproductive system tissue, breast tissue and cancer tissue, preferably breast tissue or breast cancer tissue sample. ### Claim 9 (depends on 1) The method according to claim 1, wherein said tissue sample is an estrogen receptor positive (ER+) breast cancer tissue sample. ### Claim 10 (depends on 1) The method according to claim 1, wherein said hormones are selected from the group consisting of receptor agonists or receptor antagonists, preferably estrogen receptor agonist or antagonist, progesterone receptor agonist or antagonist, androgen receptor agonist or antagonist and glucocorticoid receptor agonist or antagonist; and said drugs are selected from the group consisting of FDA-approved drugs and new drugs under clinical trials. ### Claim 11 (depends on 1) The method according to claim 1, wherein said hormones are selected from the group consisting of E2, EE2, Progesterone, R5020, DHT, dexamethasone, and progestins such as levonorgestrel, chlormadinone acetate, desogestrel, cyproterone acetate, gestodene and drospirenone, ### Claim 12 (depends on 1) The method according to claim 1, wherein said drugs are selected from the group consisting of are tamoxifen, fulvestrant, mifepristone, enzalutamide, palbociclib, abemaciclib, ribociclib, alpelisib, everolimus, olaparib, capivasertib, ER protac, AR protac and PR protac. ### Claim 13 (depends on 1) The method according to claim 1, further comprising step g) of performing one or more further assays and/or one or more further analytical techniques to determine the tissue responsiveness to hormones or drugs, preferably said further one or more assays or analytical techniques assess one or more of cell viability, cell proliferation, cell apoptosis, gene expression changes in the said tissue in response to hormones and drugs. ### Claim 14 (depends on 1) The method according to claim 1, wherein the one or more assays or analytical techniques are selected from MTT assay, Live/Dead staining, bioluminescence assay, immunofluorescent staining (IF), immunohistochemistry staining (IHC), western blot, polymerase chain reaction (PCR), RNA-seq and/or the one or more assays or analytical techniques classify the said tissue sample as responsive or non-responsive to the hormones and/or drugs. ### Claim 15 (depends on 1) The method according to claim 1, for use in hormone and drug testing for personalized medicine, drug discovery, drug development, pre-clinical or clinical studies.