Circulating tumor DNA in urothelial cancer.

Study design:

Plasma, urine and tumor specimens will be prospectively collected to determine the
longitudinal kinetics of cancer mutations in bladder cancer during chemotherapy and at
relapse. In addition, material collected in this study will be used to optimize the analysis of
ctDNA in our institute and to establish this technique at the NKI/AVL.



Study population:

The target population consists of patients who have suspected or confirmed urothelial cell
cancer of the bladder or urinary tract and who will be treated with systemic therapy. In
addition, a small number of patients with non-muscle invasive cancer will be included as
controls.



Main study parameters/endpoints:

Primary endpoint:

-The longitudinal kinetics of the spectrum of tumor mutations in the urine and plasma
of bladder cancer patients receiving systemic therapy.

-To explore the predictive value of changes in urine cfDNA and plasma ctDNA for
treatment outcome (progression free survival and pathological response rate) of
neoadjuvant chemotherapy for bladder cancer

To dermine the kinetics and spectrum of tumor mutations in the urine and plasma of bladder cancer patients receiving neoadjuvant chemotherapy for locally advanced cancer or systemic therapy for metastatic disease.

Radiological evaluation: patients will be evaluated according to local clinical practice:

During neoadjuvant treatment:

- Initial clinical staging: physical exam, TUR or bladder biopsy, FDG-PET CT
thorax/abdomen/pelvis.

-After 2 cycles to determine tumor response:
Physical exam, cystoscopy and CT
thorax/abdomen/pelvis-scan

-After 4 cycles (CT/FDG-PET)

-6 and 12 months after cystectomy

For metastatic disease: after 3 cycles of therapy by CT-scan.

Response evaluation will be assessed according to RECIST version 1.1 guidelines.25

Clinical evaluation: patients will be seen according to standard of care:

During neoadjuvant treatment before each cycle of chemotherapy to assess treatment toxicity
and response to treatment. After cystectomy every 3 months in the first year and every 6
months in the second year to assess disease control. If clinically indicated, further radiological
or metabolic response evaluation will be done.

- Peripheral blood samples

- Urine samples

- Tumor biopties