Analysis of Aqueous and Vitreous Humor

This is an open-label, pilot study of intravitreally administered ranibizumab in subjects
undergoing pars plana vitrectomy (PPV) to repair tractional retinal detachment secondary to
proliferative diabetic retinopathy, exudative serous retinal detachment, macular hole or
neovascular glaucoma.

Consented, enrolled subjects will be eligible to receive a single open-label intravitreal
injection of 0.5 mg ranibizumab administered prior to undergoing PPV if in the Retinal
Specialists opinion there could be a benefit to the disease or procedure from ranibizumab
therapy.

The proposed study will analyze aqueous humor and vitreous samples drawn at the same time
during vitrectomy to evaluate the levels of growth factors, cytokines and chemokines from
patients undergoing planned pars plana vitrectomy as part of their retinal care. For
patients receiving ranibizumab, serum samples will also be collected to assess PK. Patients
will be split into three groups based on diagnosis: Group 1: (ranibizumab) proliferative
diabetic retinopathy (tractional retinal detachment), macular degeneration with exudative or
tractional retinal detachment, macular hole or neovascular glaucoma. Group 1 may receive one
injection of intravitreal ranibizumab 0.5mg prior to vitrectomy. The injection will be
administered at least 1-14 days prior to vitrectomy. Group 2 will serve as the control group
and will not receive intravitreal ranibizumab prior to surgery.

Aqueous humor, vitreous and serum samples will be obtained to measure and compare growth
factor, cytokine, chemokine levels and ranibizumab levels (if received). Undiluted core
vitreous biopsies will be taken at the start of victrectomy. Aproximately 1 mL of undiluted
vitreous fluid will be obtained with a closed infusion line and by manual aspiration with
cutting on through the vitrectomy probe into a 2.5 mL syringe connected along the aspiration
line. The sample will be frozen immediately and stored at -80°C.

Approximately 100 microliters of aqueous humor will be collected prior to the start of
vitrectomy through anterior chamber paracentesis in the peripheral clear cornea using a 27
gauge needle on a tuberculin microsyringe from the central papillary area. Aqueous is
transferred to a vial and placed in liquid nitrogen upon removal from the anterior chamber.
Aqueous humor samples are immediately frozen at -80°C within 2 hours of collection until
transferred to outside laboratories for further analysis.

Serum samples will be obtained from patients in Group 1 at baseline (Day 0) prior to
treatment with intravitreal ranibizumab and on the day of surgery prior to the start of
vitrectomy. Serum samples will be processed according to Genentech standard operating
procedures and samples will be frozen immediately at -80C until analysis is ready to begin.

Inclusion Criteria:

- Ability to provide written informed consent and comply with study assessments for the
full duration of the study

- Age > 18 years

- Subjects with a diagnosis of tractional retinal detachment secondary to proliferative
diabetic retinopathy, age-related macular degeneration with tractional or exudative
retinal detachment, macular hole, epiretinal membrane, or neovascular glaucoma.

Exclusion Criteria:

- Subjects with history of vitrectomy in the study eye

- Presence of vitreous hemorrhage

- Subjects with history of laser photocoagulation in the study eye within 3 months of
vitrectomy

- Subjects unwilling to undergo pars plana vitrectomy

- Subjects unwilling to receive intravitreal ranibizumab prior to pars plana vitrectomy
unless diagnosis of macular hole or epiretinal membrane

- Subjects who may need or have received systemic anti-VEGF for oncology in the past
year

- Subjects with history of anti-VEGF injection within 4 months before vitrectomy