# US 2023/0407267 A1 · Uses of patient-derived scaffolds > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2023/0407267 A1 - Document ID: US20230407267A1 - Status: Published application - Kind: A1 - Application number: 18/251,326 - Filed: 2021-10-26 - Published: 2023-12-21 - Earliest claimed: 2020-11-03 - Assignees: Iscaff Pharma AB - Applicants: Iscaff Pharma AB - Inventors: Göran Landberg; Anders Ståhlberg - Web page: https://patentgrep.com/patent/US20230407267A1 ## CPC - C12N 5/0693: {Tumour cells; Cancer cells} - C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575) - C12N 2503/04: Screening or testing on artificial tissues - C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue - C12Q 2600/106: Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism - C12Q 2600/158: Expression markers - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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 ## Abstract Cell-free scaffolds derived from tumors in patients are used as in vitro cancer models and also provide information of the tumor, from which it is derived, including its susceptibility to cancer treatment. The cell-free scaffolds are also used as a predictive tool in assessing cancer treatment efficacy and in identifying immunotargets and biomarkers for cancer therapy. ## Claims ### Claim 1 A method of providing an in vitro cancer model, the method comprising: selecting (S1) a cell-free scaffold obtained from a tumor in a subject, the tumor having at least one defined tumor property and the cell-free scaffold is selected based on the at least one defined tumor property; selecting (S2) cancer cells based on the selected cell-free scaffold, wherein the selected cancer cells are not obtained from the tumor, from which the cell-free scaffold is obtained; and culturing (S3) the selected cancer cells in the selected cell-free scaffold. ### Claim 2 (depends on 1) The method according to claim 1, further comprising analyzing the expression of at least one immunomarker in the cancer cells cultured in the selected cell-free scaffold. ### Claim 3 (depends on 1, 2) The method according to claim 1 or 2, further comprising: exposing the cancer cells to a cancer treatment while culturing the cancer cells in the selected cell-free scaffold; and determining a response of the cancer cells to the cancer treatment, preferably a cancer immunotherapy. ### Claim 4 (depends on 3) The method according to claim 3, further comprising. analyzing the expression of at least one immunomarker in the cancer cells cultured in the selected cell-free scaffold prior to exposing the cancer cells to the cancer treatment; analyzing the expression of the at least one immunomarker in the cancer cells cultured in the selected cell-free scaffold after exposing the cancer cells to the cancer treatment; and comparing the expression of the at least one immunomarker in the cancer cells cultured in the selected cell-free scaffold prior to exposing the cancer cells to the cancer treatment with the expression of the at least immunomarker in the cancer cells cultured in the selected cell-free scaffold after exposing the cancer cells to the cancer treatment, wherein determining the response of the cancer cells comprises determining the response of the cancer cells to the cancer treatment based on the comparison. ### Claim 5 (depends on 3) The method according to claim 3, further comprising: determining the viability of the cancer cells cultured in the selected cell-free scaffold prior to exposing the cancer cells to the cancer treatment; determining the viability of the cancer cells cultured in the selected cell-free scaffold after exposing the cancer cells to the cancer treatment; and comparing the viability of the cancer cells cultured in the selected cell-free scaffold prior to exposing the cancer cells to the cancer treatment with the viability of the cancer cells cultured in the selected cell-free scaffold after exposing the cancer cells to the cancer treatment, wherein determining the response of the cancer cells comprises determining the response of the cancer cells to the cancer treatment based on the comparison. ### Claim 6 (depends on 3) The method according to claim 3, further comprising: determining at least one cancer cell property of viable cancer cells after exposing the cancer cells to the cancer treatment, wherein the cancer cell property is preferably selected from the group consisting of cancer stem cell-ness, proliferation, migration, apoptosis, epithelial-to-mesenchymal transition, therapy resistance, cell metabolism, cell-to-cell communication, and differentiation; and determining the response of the cancer cells comprises determining the response of the cancer cells to the cancer treatment based on the at least one determined cancer cell property. ### Claim 7 (depends on 6) The method according to claim 6, wherein determining at least one cancer cell property comprises: analyzing the expression of at least one cancer cell property marker in viable cancer cells after exposing the cancer cells to the cancer treatment; and determining the at least one cancer cell property based on the expression of the least one cancer cell property marker. ### Claim 8 (depends on 1) The method according to claim 1, further comprising determining the at least one tumor property of the tumor, from which the cell-free scaffold is obtained. ### Claim 9 (depends on 8) The method according to claim 8, wherein determining the at least one tumor property comprises determining at least one tumor property selected from the group consisting of tumor malignancy grade, cancer type, tumor location, differentiation capability, proliferation capability, infiltration capability, metastasizing capability, epithelial-mesenchymal transition (EMT) capability and cancer stem cell (CSC) capability of the tumor, from which the cell-free scaffold is obtained, selecting (S1) a cell-free scaffold obtained from a tumor in a subject, the tumor having at least one defined tumor property and the cell-free scaffold is selected based on the at least one defined tumor property; ### Claim 10 (depends on 1) The method according to claim 1, wherein selecting (S2) the cancer cells comprises selecting a cancer cell line or a primary cancer cell population based on the selected cell-free scaffold; and culturing (S3) the selected cancer cells comprises culturing cancer cells of the selected cancer cell line or the primary cancer cell population in the selected cell-free scaffold. ### Claim 11 (depends on 1) The method according to claim 1, wherein selecting (S2) the cancer cells comprises selecting cancer cells being sensitive to the influence of microenvironment of the selected cell-free scaffold. ### Claim 12 (depends on 11) The method according to claim 11, wherein selecting (S2) the cancer cell comprises selecting cancer cells having an inducible expression of at least one immunomarker, wherein expression of the at least one immunomarker is altered by the microenvironment of the selected cell-free scaffold. ### Claim 13 A method of identifying an immunotarget for cancer therapy for a subject, the method comprising: culturing (S10) cancer cells in a cell-free scaffold obtained from a tumor in the subject; analyzing (S11) expressions of immunomarkers in the cancer cells; and selecting (S12) at least one immunomarker as an immunotarget for cancer therapy for the subject based on the expressions of immunomarkers in the cancer cells. ### Claim at position 14 14-21. (canceled) ### Claim 22 A method of determining at least one tumor property of a tumor in a subject, the method comprising: culturing (S30) cancer cells, preferably breast cancer cells, in a cell-free scaffold obtained from a tumor in the subject; analyzing (S31) the expression of at least one immunomarker selected from the group consisting of programmed death ligand 1 (PDL1), colony stimulating factor 1 (CSF1) and chemokine C—C motif ligand 2 (CCL2) in the cancer cells; and determining (S32) the at least one tumor property based on the expression of the at least one immunomarker. ### Claim at position 16 23-45. (canceled) ### Claim 46 (depends on 1) The method according to claim 1, further determining susceptibility of a tumor to immunotherapy, the method comprising: selecting (S1) a cell-free scaffold obtained from a tumor in a subject, the tumor having at least one defined tumor property and the cell-free scaffold is selected based on the at least one defined tumor property; co-culturing (S40) cancer cells and leukocytes in the cell-free scaffold obtained from a tumor in a subject; determining (S42) the viability of the cancer cells; and determining (S43) the susceptibility of the tumor in the subject to immunotherapy based on the determined viability of the cancer cells. ### Claim 47 (depends on 46) The method according to claim 46, further determining at least one tumor property of a tumor in a subject, the method comprising: co-culturing (S50) cancer cells and leukocytes in the selected cell-free scaffold obtained from a tumor in a subject; analyzing (S51) the expression of at least one tumor property marker in the viable cancer cells; and determining (S52) at least one tumor property based on the expression of the at least one tumor property marker in the viable cancer cells co-cultured with the leukocytes in the cell-free scaffold, wherein determining the susceptibility of the tumor in the subject to cancer treatment is based on the determined at least one tumor property. ### Claim 48 (depends on 47) The method according to claim 47, wherein the tumor has at least one defined tumor property selected from the group consisting of tumor malignancy grade, cancer type, tumor location, differentiation capability, proliferation capability, infiltration capability, metastasizing capability, epithelial-mesenchymal transition (EMT) capability and cancer stem cell (CSC) capability. ### Claim 49 (depends on 1) The method according to claim 1, further determining susceptibility of a tumor to immunotherapy, the method comprising: selecting (S1) a cell-free scaffold obtained from a tumor in a subject, the tumor having at least one defined tumor property and the cell-free scaffold is selected based on the at least one co-culturing (S60) cancer cells and leukocytes in the cell-free scaffold obtained from a tumor in a subject; analyzing (S62) the expression of at least one immunomarker in the cancer cells from the co-culture of cancer cells and leukocytes in the cell-free scaffold; and determining (S63) the susceptibility of the tumor in the subject to immunotherapy based on the expression of the at least one immunomarker. ### Claim 50 (depends on 49) The method according to claim 49, further comprising: culturing cancer cells in a cell-free scaffold obtained from a tumor in the subject; analyzing the expression of the at least one immunomarker in the cancer cells from the culture of cancer cells in the cell-free scaffold; and comparing the expression of the at least one immunomarker in the cancer cells from the culture with the expression of the at least one immunomarker in the cancer cells from the co-culture, wherein determining (S63) the susceptibility comprises determining the susceptibility of the tumor in the subject to immunotherapy based on the comparison. ### Claim 51 (depends on 49) The method according to claim 49, further comprising determining at least one tumor property of viable cancer cells co-cultured with the leukocytes in the cell-free scaffold, wherein determining (S63) the susceptibility comprises determining the susceptibility of the tumor in the subject to immunotherapy based on the expression of the at least one immunomarker and based on the at least one tumor property.