Microtentacle imaging in patient tumor samples

Inventors

Martin, Stuart S.Jewell, ChristopherAndorko, JamesSooklal, ElisabethWhipple Bettes, RebeccaChakrabarti, Kristi

Assignees

University of Maryland BaltimoreUniversity of Maryland College Park

Publication Number

US-12146874-B2

Publication Date

2024-11-19

Expiration Date

2035-04-03

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Abstract

The present invention provides a method for imaging microtentacles on isolated, living, non-adherent primary tumor cells from a cancer subject comprising: i) obtaining one or more living, non-adherent primary tumor cells that has been isolated from a solid tumor from the subject; and ii) imaging the one or more living, non-adherent primary tumor cells and detecting the microtentacles.

Core Innovation

The invention provides a method for imaging microtentacles on isolated, living, non-adherent primary tumor cells taken directly from a cancer patient's solid tumor. The process comprises obtaining one or more living, non-adherent primary tumor cells that have been isolated by dissociation ex vivo from the primary solid tumor and imaging these cells to detect and analyze the presence of microtentacles. This allows for assessment of functional tumor cell phenotypes immediately after isolation.

The central problem addressed is the inadequacy of current cancer prognostic and treatment strategies in identifying patients at risk for metastasis, as circulating tumor cells (CTCs) are often present in low numbers and standard imaging or molecular tests may not reveal their metastatic potential. Additionally, existing cancer drug development and clinical assessment largely overlook the effects of drugs on non-adherent, free-floating tumor cells, which are key players in metastatic spread.

This invention enables direct visualization and scoring of microtentacles in freshly isolated tumor cells, which serve as indicators of metastatic and stem cell potential. The approach also includes determining the response of these cells to candidate drugs by measuring changes in microtentacle incidence, thereby assisting in the selection of therapies that inhibit metastatic capability and optimizing individualized patient treatment.

Claims Coverage

The independent claims of this patent cover four main inventive features related to methods for imaging and analyzing microtentacles in live primary tumor cells for prognosis, therapy guidance, drug screening, and stem cell potential determination.

Method for imaging microtentacles on dissociated live primary tumor cells

A method comprising: 1. Obtaining one or more live primary tumor cells isolated by ex vivo dissociation from a primary solid tumor in a cancer patient. 2. Imaging the live primary tumor cells and detecting the presence of microtentacles. This method is used to provide prognostic information and guide the patient's treatment.

Method for identifying metastatic risk via microtentacle scoring

A method comprising: 1. Obtaining one or more live primary tumor cells isolated by dissociation ex vivo from a primary solid tumor in a subject. 2. Imaging the live primary tumor cells. 3. Scoring the imaged cells for microtentacles to determine whether the subject has an increased likelihood of having or developing metastatic cancer. The scoring can include determining whether cells have two or more protrusions longer than the cell radius.

Method for determining drug effects on microtentacle formation or stability

A method comprising: 1. Obtaining one or more live primary tumor cells from a solid tumor by ex vivo dissociation. 2. Contacting the cells with a candidate drug. 3. Imaging the treated cells. 4. Scoring the imaged cells for microtentacles to determine if the drug inhibits or promotes microtentacle formation and/or stability. Optionally includes comparison to untreated cells and directions for using these results to guide cancer therapy.

Method for determining stem cell potential based on microtentacle analysis

A method comprising: 1. Obtaining one or more live primary tumor cells from a solid tumor by dissociation ex vivo. 2. Imaging the live primary tumor cells. 3. Scoring the imaged cells for microtentacles to determine the stem cell potential of the tumor cell. Cells with two or more protrusions longer than the cell radius are considered to have greater stem cell potential.

The inventive features encompass methods for directly imaging and analyzing live, dissociated primary tumor cells for the presence of microtentacles, using this information to assess metastatic risk, guide drug selection, and determine stem cell potential.

Stated Advantages

Provides an immediate functional phenotype measurement of tumor cell metastatic and stem cell potential without reliance on gene or protein expression profiles.

Enables rapid testing using very few cells, avoiding long-term cell culturing and reducing the risk of altering tumor cell properties compared to the patient's original tumor.

Allows real-time assessment of drug effects on microtentacles, enabling quick guidance for personalized therapy selection.

Directly assesses tumor cell response to candidate drugs prior to treatment, potentially improving prognosis and treatment efficacy.

May improve identification of patients at higher risk of metastasis by evaluating microtentacle formation in cells from tumors or biopsies.

Documented Applications

Imaging and analysis of microtentacles in primary tumor cells to provide prognostic information regarding metastatic potential in cancer patients.

Screening and selection of cancer drugs for individual patients by determining their effects on microtentacle formation and stability in patient-derived tumor cells.

Assessing and predicting stem cell potential in tumor cells based on microtentacle presence and characteristics.

Application to a broad range of tumor types including breast, prostate, lung, bladder, pancreatic, brain, liver, testicular, thyroid, skin, colon, ovarian, cervical, and uterine cancers.

Use in medical devices, including microfluidic slides and patterned substrates, for high-throughput imaging and drug screening of live patient tumor cells.

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