Initial Evaluation of a Telomerase-based Circulating Tumor Cell Assay in Bladder Cancer Cohorts

Inclusion Criteria:

- a.Urothelial carcinoma or mixed urothelial carcinoma arising from the bladder

- b.Age 18 or older

- Inclusions by cohort Cohort 1. Patients with localized muscle-invasive disease
(cT1,cN0, M0) at the time of initial diagnosis or who have progressed from
superficial disease (cT2) who have not had any systemic chemotherapy and who will be
treated with upfront radical cystectomy. PriorTURBT and intravesical therapies are
allowed but should not have been done within one week of CTC measurement. If the
patient is found to have no evidence of tumor (pT0) upon evaluation of the cystectomy
specimen, the patient will not be dropped. If the patient is found to have evidence
of progression by any clinical evaluation performed within 2 months of the
post-treatment sample, the patient will be dropped from Cohort 1 and included in
Cohort 9 instead.

- Cohort 2. Patients with localized muscle-invasive disease (cT1, cN0, M0) at the time
of initial diagnosis prior to definitivechemo-radiation. Prior TURBT and intravesical
therapies are allowed but should not have been done within one week of CTC
measurement. If the patient is found to have evidence of progression by anyclinical
evaluation performed within 2 months of the post-treatment sample, the patient will
be droppedfrom Cohort 2 and included in Cohort 9 instead.

- Cohort 3. Patients with localized muscle-invasive disease (cT1, cN0, M0) at the time
of initial diagnosis or who have progressed from superficial disease (cT2) who have
not had any systemic chemotherapy and who will be treated with neoadjuvant
chemotherapy. Prior TURBT and intravesical therapies are allowed but should not have
been done within one week of CTC measurement. If the patient is found to have
evidence of progression by any clinical evaluation performed within 2 months of the
post-treatment sample, the patient will be dropped from Cohort 3 and included in
Cohort 9 instead.

- Cohort 4. Patients with metastatic disease (anyT, anyN, M1) at initial presentation
or recurrent after definitive treatment prior to any salvage systemic chemotherapy.
If the patient is found to have evidence of progression by any clinical evaluation
performed within 2 months of the post-treatment sample, the patient will be dropped
from Cohort 4 and included in Cohort 9 instead.

- Cohort 5. Patients with newly diagnosed non-muscle-invasive bladder cancer (T2) prior
to any intravesical therapy. If the patient is found to have evidence of progression
by any clinical evaluation performed within 2 months of the post-treatment sample,
the patient will be dropped.

- Cohort 6. Patients with muscle-invasive bladder cancer after radical cystectomy with
or without peri-operative systemic chemotherapy (pT2, anyN, M0) who will be treated
with adjuvant radiation. If the patient is found to have evidence of progression by
any clinical evaluation performed within 2 months of the post-treatment sample, the
patient will be dropped from Cohort 6 and includedin Cohort 9 instead.

- Cohort 7. Patients with a history of muscle invasive bladder cancer (cT1, any N,M0)
who have no evidence of disease at least 2 years after the date of radical
cystectomy. If the patient is found to have evidence of progression by any clinical
evaluation performed within 2 months of the post-treatment sample, the patient will
be dropped from Cohort 7 but may be considered for inclusion in Cohort 4.

- Cohort 8. Patients with a history of muscle invasive bladder cancer (cT1, any N, M0)
who have no evidence of disease at least 2 years after the completion of definitive
chemo-radiation. If the patient is found to have evidence of progression by any
clinical evaluation performed within 2 months of the post-treatment sample, the
patient will be dropped from Cohort 8 but may be considered for inclusion in Cohort
4.

- Cohort 9. Patients who were initially included in Cohorts 1-6 but were found to have
progression at the time of their post-treatment blood draw. Of note, patients may be
included in more than one cohort over time so long as inclusion and exclusion
criteria are met. In addition, twenty samples from normal controls without any
history of malignancy will be used for this study but havealready been collected as
part of a separate protocol.

Exclusion Criteria:

- pure squamous or pure adenocarcinoma bladder cancer histology. This exclusion may be
waved at the discretion of the protocol PI. 2. Prior cancer diagnosed within 3 years
of enrollment other than a prior bladder, prostate, or skin cancer. This exclusion
may be waved at the discretion of the protocol PI.