Methods of detecting bladder cancer
Inventors
McCann, Leena • Wallace, Stacey Ellen • Lai, Edwin Wei-Lung • Higuchi, Russell
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Assignees
Member
CepheidCepheid is a global leader in molecular diagnostics, dedicated to improving healthcare by developing, manufacturing, and marketing automated, easy-to-use molecular systems and tests. Their mission is to provide rapid, accurate, and actionable genetic testing for a wide range of infectious diseases, oncology, and human genetics. Cepheid's flagship GeneXpert System delivers scalable, sample-to-answer PCR testing for institutions of any size, supporting both centralized and decentralized care. The company is committed to expanding access to high-quality diagnostics worldwide, supporting public health initiatives, driving innovation in molecular testing, and advancing sustainability and responsible business practices.
Cepheid is a global leader in molecular diagnostics, dedicated to improving healthcare by developing, manufacturing, and marketing automated, easy-to-use molecular systems and tests. Their mission is to provide rapid, accurate, and actionable genetic testing for a wide range of infectious diseases, oncology, and human genetics. Cepheid's flagship GeneXpert System delivers scalable, sample-to-answer PCR testing for institutions of any size, supporting both centralized and decentralized care. The company is committed to expanding access to high-quality diagnostics worldwide, supporting public health initiatives, driving innovation in molecular testing, and advancing sustainability and responsible business practices.
Publication Number
US-12000001-B2
Publication Date
2024-06-04
Expiration Date
Abstract
Compositions and methods for detecting bladder cancer are provided. In some embodiments, methods of detecting low grade bladder cancer are provided. In some embodiments, methods of monitoring recurrence of bladder cancer are provided. In some embodiments, the methods comprise detecting androgen receptor (AR) and/or uroplakin 1B (UPK1B).
Core Innovation
Compositions and methods for detecting bladder cancer are provided. In particular, bladder cancer markers and panels of markers useful in the detection of bladder cancer are provided.
Improved methods for early detection of bladder cancer are needed. An accurate urine-based diagnostic test that does not rely on cytology could reduce the need for costly and invasive cystoscopy and labor-intensive and potentially subjective cytology assays.
In some embodiments, the levels of androgen receptor (AR) and/or uroplakin 1B (UPK1B) mRNA, and optionally, at least one mRNA selected from CRH, IGF2, KRT20 and ANXA10 mRNA, are measured, for example, by quantitative RT-PCR, and the results can be used to determine whether or not a subject has bladder cancer. In some embodiments, the sample is a urine sample and the present methods are used to monitor subjects with a history of bladder cancer for tumor recurrence or to detect bladder cancer in subjects with no history of bladder cancer.
The present assays do not rely on cytology and instead rely on the polymerase chain reaction (PCR), and can be carried out in a substantially automated manner, for example, using the GeneXpert® system. The present assays can be completed in under 3 hours, and in some embodiments, under 2 hours, and can be carried out on much smaller volumes of urine (in some embodiments, 5 ml or less), allowing for a fast, one-pot reaction for diagnosis of bladder cancer, which in many instances can be carried out at the point of care.
Claims Coverage
The independent claim is claim 1 and recites a method of monitoring anti-androgen therapy response in a human subject with bladder cancer. The claim contains six main inventive features.
Monitoring anti-androgen therapy response by comparing marker levels at two time points
A method comprising detecting the levels of each marker of a set of three to six bladder cancer markers in a sample from the subject at a first time point and at a second time point that is after the first time point, and comparing the levels at the two time points, wherein the subject is undergoing anti-androgen therapy at the second time point.
Set of bladder cancer markers comprising UPK1B and IGF2 and at least one additional marker
The set of bladder cancer markers comprises uroplakin 1B (UPK1B) and insulin-like growth factor 2 (IGF2) and at least one marker selected from androgen receptor (AR), corticotrophin releasing hormone (CRH), keratin 20 (KRT20), and annexin 10 (ANXA10).
Detection by contacting RNA with primer pairs and forming amplicons and probes
Detecting comprises contacting RNA from the sample with a set of bladder cancer marker primer pairs comprising a primer pair for detecting UPK1B and a primer pair for detecting IGF2, forming a set of bladder cancer marker amplicons, and contacting the bladder cancer marker amplicons with a set of bladder cancer marker probes comprising a probe for detecting the UPK1B amplicon and a probe for detecting the IGF2 amplicon.
IGF2 probe comprising at least 8 contiguous nucleotides of SEQ ID NO: 34 and probe length limitation
The probe for detecting the IGF2 amplicon comprises at least 8 contiguous nucleotides of SEQ ID NO: 34, and each probe is less than 30 nucleotides long.
Decrease in marker level indicates response to anti-androgen therapy
A decrease in the level of at least one marker in the set of bladder cancer markers at the second time point relative to the first time point indicates that the subject is responding to anti-androgen therapy.
Primer and amplicon constraints and RT-PCR detection
The claim specifies primer/probe constraints and detecting steps; dependent claims recite that detecting comprises RT-PCR, that a Ct value or a ΔCt value can be compared to a threshold, that each bladder cancer marker primer pair produces an amplicon that is 50 to 500 nucleotides long, and that at least one primer pair spans an intron.
Claim 1 covers a method combining serial measurement of a specified marker panel that includes UPK1B and IGF2 (with optional markers AR, CRH, KRT20, ANXA10), nucleic acid amplification-based detection using specified primer and probe constraints (including an IGF2 probe comprising SEQ ID NO: 34), and interpreting a decrease in marker level between two time points as an indication of response to anti-androgen therapy.
Stated Advantages
The present assays do not rely on cytology and instead rely on PCR, avoiding cytology's requirement for trained cytologists.
The assays can be carried out in a substantially automated manner, for example, using the GeneXpert® system.
The present assays can be completed in under 3 hours, and in some embodiments, under 2 hours.
The present assays can be carried out on much smaller volumes of urine (in some embodiments, 5 ml or less).
An accurate urine-based diagnostic test that does not rely on cytology could reduce the need for costly and invasive cystoscopy and labor-intensive and potentially subjective cytology assays.
Documented Applications
Detecting bladder cancer.
Detecting low grade bladder cancer.
Monitoring recurrence of bladder cancer.
Monitoring anti-androgen therapy in a subject with bladder cancer by serial measurement of bladder cancer markers.
Detecting marker mRNAs in urine samples and bladder washing samples, including detection in urothelial cells.
Point-of-care testing using an automated system such as the GeneXpert® system.
Screening healthy individuals and asymptomatic individuals with risk factors for bladder cancer.
Assessing the effectiveness of a treatment for bladder cancer and monitoring target RNA levels at regular or semi-regular intervals.
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