photodynamic therapy of intraocular tumors examination of...

(CANCER RESEARCH 45, 3718-3725, August 1985]

Photodynamic Therapy of Intraocular Tumors Examination of HematoporphyrinDerivative Distribution and Long-Term Damage in Rabbit Ocular Tissue1

Charles J. Gomer,2 James V. Jester, Nicholas J. Razum, Bernard C. Szirth, and A. Linn Murphree

Clayton Center lor Ocular Oncology, Childrens Hospital of Los Angeles [C. J. G., N. J. R., B. C. S., A. L. M.j, and Departments of Pediatrics (Division of Hematology-Oncology) [C. J. G.Â¡,Radiation Oncology (C. J. G.Â¡,and Ophthalmology [J. V. J., A. L M.], USC School oÃMedicine, Los Angeles, California 90027

ABSTRACT

Studies were performed to determine the distribution of he-

matoporphyrin derivative (HPD) in ocular structures and to characterize long-term damage associated with ocular HPD photo-

dynamic therapy. Pigmented rabbits with an amelanotic melanoma heterotransplanted to the iris were used to obtain quantitative tissue levels of HPD as well as to document HPD localization by fluorescence microscopy. HPD was administered i.V.,and tissue concentrations of HPD were determined by spectro-

fluorometry following porphyrin extraction. Vascular structuressuch as the tumor, iris, and choroid-retina as well as the aqueous

fluid from eyes containing tumors demonstrated rapid HPD localization. The sclera had minimal HPD uptake, and the drugwas not detected in avascular structures such as the lens orcornea. HPD was cleared from all ocular structures except thetumor and choroid-retina by 24 h following injection. Fluores

cence microscopy data indicate that HPD remained in the avascular photoreceptor cell outer segments of the retina. Long-termdamage was documented in rabbits which received HPD photody-namic therapy to a 1-sq cm area of retina via transpupillary lightdelivery. Acute damage to the exposed area of retina (in theform of a chorioretinal scar) could be induced. This damage waspermanent but not progressive. Lens opacities were not observed, and the cornea, aqueous, and vitreous remained clearon all test eyes. The results from these studies suggest thatHPD photodynamic therapy may provide a selective and safeapproach to the treatment of ocular tumors.

INTRODUCTION

Clinical treatment of solid tumors by HPD3 PDT continues to

show promise (1, 2). HPD exhibits properties of both preferentialtumor localization and visible light-activated tumor destruction

via photosensitization (3, 4). A general treatment protocol forPDT consists of i.v. administration of HPD followed 2 to 3 dayslater by localized exposure of the tumor to visible light. Red lightis most often utilized in PDT due to the superior tissue penetration characteristics of this wavelength (5), and the tumor destruction following HPD PDT is believed to be caused by singletoxygen ('Oa) which is produced via a type II photochemical

reaction (4). Advances in the development of laser systems and

1This investigation was performed in conjunction with the Clayton Foundationfor Research and was supported in part by USPHS Grants CA-31230 and CA-31885 awarded by the National Institutes of Health, Department of Health andHuman Services.

2To whom requests for reprints should be addressed, at Clayton Center for

Ocular Oncology. Childrens Hospital of Los Angeles, 4650 Sunset Boulevard, LosAngeles. CA 90027

3The abbreviations used are: HPD, hematoporphyrin derivative; PDT, photody

namic therapy.Received 1/2/85; revised 3/25/85; accepted 4/17/85.

procedures for light delivery, as well as continued preclinicalresearch, have led to the utilization of HPD PDT for the treatmentof numerous types of malignant tumors (6-14). Several current

reviews describe the preclinical and clinical HPD investigationsperformed over the past 5 to 7 years (1,2,15,16).

A recent application of HPD PDT is for the treatment ofintraocular tumors such as uveal melanoma and retinoblastoma(7, 14, 17). The accessibility of ocular tumors and the opticalproperties of the eye appear to be compatible with HPD PDT.Depending on the clinical situation, most ocular tumors arecurrently treated by combinations of periodic observation, cryo-therapy, photocoagulation, radioactive plaques, external beamradiation therapy, or enucleation (18-21). Unfortunately, each of

these clinical modalities has various limitations or disadvantageswhich suggest that new types of therapy should be investigated.Cryotherapy and photocoagulation are effective only in the treatment of small lesions (21). Retinoblastoma is considered a radiation-sensitive tumor, but recent surveys report up to 50% failure

rates following external beam radiation for unilateral and bilaterallesions (22). In addition, there is evidence that radiotherapy forbilateral retinoblastoma may play a role in the appearance ofsecond primary tumors (22). Brachytherapy using 60Co plaques

is effective in destroying choroidal melanoma, but failure rates ofapproximately 30% are reported (23), and intraocular complications frequently occur (24). Charged particles (protons and heliumions) offer improved dose distributions and possibly enhancedbiological advantages for the treatment of choroid melanomas(25, 26). However, the expense necessary to build, maintain,and operate cyclotrons suggests that widespread use of thesemodalities will be difficult to achieve. Since the size and placement of ocular tumors are often limiting factors in the effectiveness of current therapies, it is necessary to explore new methodsof treating primary and recurrent tumors of the eye.

HPD-induced fluorescence is observed in transplanted anterior

chamber tumors in the hamster and rabbit (27). In addition,porphyrin-induced fluorescence is observed in an amelanotic

melanoma transplanted in the rabbit choroid following HPDadministration (28). Human retinoblastoma heterotransplanted tonude mouse eyes retains [3H]HPD, and this tumor model can be

destroyed by HPD PDT (29, 30). Acute normal ocular tissuedamage can be induced in rabbits by HPD PDT (30,31). Damagein the form of retinal edema, detachment, and necrosis is seenwithin 2 days of ocular PDT, but the area of damage is limitedto the treatment field in all but the highest doses of PDT. Theuse of HPD PDT in treating anterior chamber tumors in rabbitshas been described by several groups (32-34). In most cases, a

rapid alteration in the appearance of the tumor appears within24 h of treatment and is characterized by blanching, edema, andhemorrhage. Anterior chamber amelanotic melanomas measuring 4 to 5 mm in diameter and 2 to 3 mm high can be completely

CANCER RESEARCH VOL. 45 AUGUST 1985

3718

Research. on January 12, 2020. © 1985 American Association for Cancercancerres.aacrjournals.org Downloaded from

http://cancerres.aacrjournals.org/

HPD UPTAKE AND PHOTOSENSITIZATION IN THE EYE

eradicated using a HPD PDT protocol consisting of HPD (5 mg/kg) and 90- to 180-J/sq cm dose of 630 nm light delivered at a

power density of 150 milliwatts/sq cm (32).HPD PDT has been used clinically to treat select cases of

uveal melanoma and retinoblastoma (7,14). Most ocular tumorsrespond to PDT by exhibiting posttreatment hemorrhage andblanching, but long-term follow-up of these patients is not yet

available. It is clear, however, that additional preclinical studiesare needed prior to the initiation of meaningful clinical trials. Theexperiments described in this paper were designed to documentHPD distribution in ocular structures and to characterize thelong-term toxicity effects of ocular HPD PDT.

MATERIALS AND METHODS

Drugs. HPD was obtained from Photofrin Medical, Inc., Cheektowaga,NY, and was kept at 4Â°Cin the dark until used. A solution of ketamine

(30 mg/kg), acepromazine 93 mg/kg), and atropine sulfate (0.15 mg/kg)was administered i.m. to anesthetize animals prior to all experimentalprocedures. A solution of 1% cyclopentolate and 10% phenylephrinehydrochloride was used to dilate pupils and i.v. sodium pentobarbital(120 mg/kg) was used for euthanasia.

Animal and Tumor Models. Pigmented rabbits weighing approximately 2 kg were obtained from ABC Babbitry, Pomona, CA, and wereentered into studies 1 week after delivery. The amelanotic hamstermelanoma heterotransplanted onto the iris of the rabbit eye was used inall HPD tissue distribution studies. Single-nodule tumors were obtainedby placing a 1 mm3 piece of viable tumor (obtained from a stock tumor

growing in the anterior chamber of a donor rabbit) onto the iris througha paracentesis incision at the limbus. The growth rate of the lesion isapproximately 1 mm in diameter per day (31). Studies which involvedtumors were initiated when the lesions were between 5 and 8 mm indiameter (approximately 10 days following transplant).

Light Source and Delivery Systems. All HPD PDT experiments wereperformed with a dye laser (using Kilon red dye) pumped by a 5-watt

argon ion laser (Spectra Physics, Mountain View, CA). The outputwavelength from the dye laser was set at 630 nm and checked with ascanning monochromator (American ISA, Inc., Metuchen, NJ), and thelight power density was measured with a thermopile (Model 210; Coherent Radiation, Palo Alto, CA). Light from the laser was interfaced via aspatial filter (Spectra Physics) through a single-stop quartz fiber optic

bundle. A microlens was fitted on the distal end of the fiber optic forproduction of a uniform light treatment field (34).

An 18-watt krypton ion laser running at 407 to 413 nm (CoherentRadiation) was used to visualize porphyrin-induced fluorescence in tissue

precipitates.HPD Distribution in Ocular Tissue. Rabbits with amelanotic mela

noma growing on the iris of one eye received an i.v. injection of HPD ata dose of either 5 mg/kg for quantitative uptake studies or 10 mg/kg forfluorescence microscopy studies. At selected time intervals followinginjection, the animals were sacrificed, and ocular structures and fluids(cornea, aqueous, tumor, iris, lens, vitreous, retina-choroid, and sclera)

were collected. The retina and choroid from each test eye had to becombined in order to obtain enough material for porphyrin determinations.Specimens were either assayed for quantitative porphyrin content usinga tissue extraction procedure or examined by fluorescence microscopy.

Tissue samples for quantitative analysis of porphyrin levels were firstlyophilized. The dried tissues were then ground into a powder, and 20-mg samples (5-mg samples for iris and choroid-retina) were subjected to

porphyrin extraction using a modification of an acid extraction procedure(36). Specimens were homogenized with TenBroeck tissue grinders in a7-ml solution consisting of 1% sodium dodecyl sulfate in 1 N perchloric

acid:methanol (1:1, v/v). The homogenates were centrifugea at 1000rpm for 10 min, and the supernatants were collected. Tissue precipitates

were then reextracted, and resulting supernatants were combined. Theremaining tissue precipitates were examined for porphyrin-induced fluorescence using the 407 to 413 nm lines of a krypton laser. Porphyrin-

induced fluorescence was not observed in the tissue precipitates following the double extraction procedure.

Porphyrin levels in the supernatants were quantified by fluorescenceusing a scanning spectrofluorometer (SLM Instruments, Ine, Urbana, IL).The solutions were excited at 405 nm, and emission scans from 550 to700 nm were obtained. Fluorescence values at 600 nm (emission maximum for HPD in extraction medium) were recorded for each tissuesample, and background fluorescence (from control samples) was subtracted from these values. Porphyrin levels were then determined fromcalibration curves which were produced by adding known amounts ofHPD to control samples of each tissue prior to porphyrin extraction.Aqueous and vitreous levels of porphyrins were determined directly byfluorescence (without extraction) and then compared to calibrationcurves. Serum levels of porphyrin were determined by spectrophotom-

etry.At the time of sacrifice, tissue specimens to be examined by fluores

cence microscopy were embedded in O.C.T. compound (Lab-Tek; Miles

Laboratories) and frozen in liquid nitrogen. Sections (10 ^m) were cutusing a cryostat and mounted on glass slides with glycerol:phosphate-buffered saline (1:2, v/v) containing p-phenylenediamine (1 mg/ml) as a

quenching agent. Specimens were examined under phase contrast andfluorescence using a Zeiss fluorescence microscope with 390 to 440 nmand 405 nm interference (exciter) filters, a 580 nm chromatic beamsplitter, and a 590 nm barrier filter.

Chronic Ocular Tissue Damage. The procedure utilized to examinelong-term damage following HPD PDT to normal rabbit eyes was similar

to the method previously used in examining acute damage (31, 32).Briefly, normal pigmented rabbits were given an i.v. injection of HPD(2.5, 5, 10, or 20 mg/kg), and then 24 or 48 h later a 1-sq cm area of

the fundus of each test eye was irradiated with red light (630 nm) at adose rate of 150 milliwatts/sq cm. Total light dosages ranged from 36to 180 J/sq cm. A total of 24 eyes from 12 rabbits were treated andanalyzed for long-term toxicity. Ocular toxicity was documented by serialfundus photography, fluorescein angiography, and slit-lamp examinationduring the follow-up period (9 to 30 months). An Equator Plus Cameraequipped with direct illumination and a 148-degree wide-angle lens was

used for both fundus photography and fluorescein angiography. Serialexamination of the rabbit lens was performed with a Kowa hand-held slit

lamp. Following the observation period, the test animals were sacrificed,and the test eyes were fixed in 10% buffered formalin, embedded inparaffin, sectioned, and stained with hematoxylin and eosin.

RESULTS

The distribution of HPD in ocular structures and fluids is shownin Table 1. Vascular structures such as the iris, choroid-retina,

and tumor all demonstrated rapid localization of HPD. In addition,the aqueous in eyes with anterior chamber tumors also accumulated significant levels of HPD soon after drug administration.HPD was not detected in avascular structures such as the corneaor lens. The sclera and aqueous (from eyes without tumors) hadextremely low concentrations of HPD at early time intervals afterdrug administration. At extended time periods following injection(24 h), HPD was cleared from the iris, sclera, and aqueous. HPDlevels remained high in the tumor and choroid-retina at both 24

and 48 h.Phase-contrast and fluorescence micrographs of ocular struc

tures were obtained at various time intervals following HPDadministration. Micrographs of tumor tissue at 6 h following druginjection are shown in Fig. 1. While HPD was present in theentire tumor, the highest intensity of fluorescence was observed


3719




Table 1Ocular distribution ol HPD (5-mg/kg dose) in rabbits

Time post-injection

(h)0.250.51232448HPD

tissue concentrations (^g/g dry wt orng/ml)Serum84

Â±9.5"(4)c60

Â±11.0(4)45

Â±3.4(4)35.5

Â±6.4(4)27.0

Â±5.5(4)5.13

Â±0.05(3)2.7

Â±0.6(3)Tumor5.34

Â±1.98(5)3.14

Â±1.61(6)4.4

Â±2.69(4)4.75

Â±1.27(3)5.19

Â±1.80(4)7.2

Â±3.26(6)5.22

Â±2.23(6)Cornea0(3)0(3)0(4)0(3)0(3)0(6)0(2)Iris16.6

+10.02(4)14.99(2)13.67

Â±9.75(3)7.58

Â±5.36(3)11.91

Â±4.11(4)1.27

Â±0.3(7)0.37

Â±0.2(5)Lens0(6)0(6)0(6)0(7)0(4)(10)0(5)Choroid

andretina24.1

2 Â±15.95(5)11.12

Â±2.85(5)21

.53Â±6.61(3)5.9

Â±3.82(4)12.31

Â±4.16(6)6.71

Â±1.67(4)6.41

Â±1.75(4)Sclera0.01

Â±0.01(4)0.17

Â±0.1(8)0.25

Â±0.16(5)0.79

Â±0.28(7)0.16

Â±0.01(4)0(5)0(6)Aqueous0

(tumor)0.25

Â±0.17(3)1.25

Â±0.78(2)3.7

Â±1.27(3)4.1

5Â±1.65(2)7.55

Â±5.28(3)0.25

Â±0.21(3)0.1

2Â±0.02(5)Aqueous0.01

Â±0.02(3)0.31

Â±0.23(4)0.27

Â±0.22(4)0.34

Â±0.17(4)0.23

Â±0.06(7)0.23

Â±0.13(4)0.01

Â±0.01(2)Vitreous0(4)0(3)0(4)0(3)0(2)0(4)0(3)

" Aqueous from eye which contained a tumor.0 Mean Â±SD.c Numbers in parentheses, number of samples analyzed (1 sample/rabbit).

in what appeared to be necrotic areas of the tumor mass (Fig.1, arrow). HPD was not observed to localize in or around clearlyidentifiable vascular channels (Fig. 1, open arrow).

Fluorescence localization patterns of HPD in ocular structuresare shown in Figs. 2 to 4. Figs. 2 and 3 illustrate the HPDdistribution in the ciliary body (Fig. 2) and the choroid-retina (Fig.

3) at 30 min following injection. At this time point, HPD fluorescence was localized primarily to the small cells or capillaries inthe ciliary processes (Fig. 2, small arrows) and to the ciliaryepithelium (Fig. 2, arrowhead). In addition, HPD was observed inthe choroid and retinal pigment epithelium (Fig. 3). Minimalfluorescence was seen in the photoreceptor cell outer segmentsof the retina at this time period, and fluorescence was notobserved in the sclera. Fig. 4 shows the HPD distribution in thechoroid-retina at 24 h following drug injection. At this time period,

HPD fluorescence was observed in the retinal epithelium and theretinal photoreceptor cell outer segments.

Table 2 summarizes the effects of HPD PDT which relate tolong-term ocular damage. The length of follow-up ranged from 9

to 30 months. Damage to the exposed area of retina wasobserved and documented using the techniques of fundus photography, fluorescein angiography, and histopathological examination. The retinal damage induced by HPD PDT was a chorio-retinal scar. Slit-lamp examinations demonstrated normal suture

lines in the lenses of all test eyes, but lens opacities were notobserved. In addition, the cornea, aqueous, and vitreous of alltest eyes remained clear throughout the follow-up period.

Fig. 5 illustrates typical fundus photographs of Rabbit Eye 5from the long-term chronic damage study. Fig. 5A shows the

rabbit retina prior to treatment. The area of acute chorioretinaldamage observed 2 days following HPD PDT (10 mg/kg, 90 J/sq cm) is shown in Fig. 56. The damaged area appeared similarto that observed previously in acute toxicity experiments (30),and this would suggest that retinal detachment as well asnecrosis was induced. Fig. 5, C and D documents the appearanceof the treatment area at 7 days and 6 months, respectively. Thechorioretinal scar was present 21 months following treatment asshown in Fig. 5E. Retina outside the treatment field appearednormal as determined by fundus photography, fluorescein angiography, and histopathological examination.

Table 2Chronic normal ocular tissue damage following HPDPDT

Eye123456789101112131415161718192021222324HPD

dose8(mg/kg)20201010101010101010555555552.52.52.52.500Lightdose"

(J/sqcm)18018018018090909036363690909090363636369090363600Follow-uptime(mo)1616161625239252393022161030221610232223222424OpacitiesFunriiisappearance0Lens0Corneal"++++++

â€”â€”+++â€”â€”+++-â€”+++-â€”+++â€”â€”+++â€”â€”+++-â€”++â€”â€”+++â€”-+++â€”â€”++â€”â€”++â€”â€”+++â€”â€”++â€”â€”___+â€”â€”+++â€”â€”___-t-â€”â€”___+â€”â€”____

Administration i.v. 48 h prior to PDT.6 Light (630 nm) delivered at 150 milliwatts/sq cm.0 Fundusappearance:+++, exudative retinaldetachment and chorioretinalscar

in treatment field; ++, focal area of exudative retinal detachment and chorioretinalscar; +, minimalexudative retinal detachment; -, normal.

" Lens and corneal appearance,â€”,normal.

DISCUSSION

Localization of HPD in ocular structures and characterizationof long-term ocular toxicity following HPD PDT have been ob

tained using a rabbit model. Current experiments were designedso that a clinically relevant dose of HPD (5 mg/kg) was utilizedin quantitative HPD localization experiments. Relatively highdoses of HPD (20 mg/kg) were included in chronic toxicityexperiments in order to document the types and severity ofocular toxicity which could be induced by HPD PDT underextreme parameters. The amelanotic melanoma can be hetero-transplanted to either the choroid or iris of rabbits. While thechoroidal placement is a more realistic anatomical location (mosthuman ocular tumors are choroidal), we utilized experimental iris


3720




tumors to ensure that a standard tumor size was examined inall HPD uptake studies.

The pharmacokinetics of HPD has been examined previouslyusing radiolabeled compounds ([14C] and [3H]HPD) or by spec-trofluorometric techniques. The use of [14C]HPD in large animalsis cost prohibitive, and [3H]HPD can degrade to produce radio

active contaminants (37). Therefore, the uptake and distributionof HPD in ocular structures were documented using a spectro-

fluorometric procedure following porphyrin extraction from tissue. The efficiency of any drug extraction techniques may be apotential variable. In our experiments, however, the tissue precipitates which were obtained following extraction did not containdetectable levels of porphyrin fluorescence. While this indicatesthat a high percentage of HPD was extracted from the tissuespecimens, it does not rule out the possibility that HPD-induced

fluorecence may be quenched by pigmented tissue precipitatesof iris or choroid. Fluorescence microscopy data correlated withthe quantitative spectrofluorometric data with respect to documenting which ocular structures accumulated HPD. Results fromour study indicate that HPD was retained in an experimentalocular melanoma. These findings confirm an earlier investigationdescribing HPD-induced fluorescence in the same tumor when itis transplanted to the choroid of the rabbit eye (28). The highlevels of HPD in the aqueous of tumor-containing eyes indicated

that porphyrin leaks from incompetent vessels of the tumor andthat alterations to the blood-aqueous barrier may be induced.

The concentrations of HPD found in the experimental oculartumor may be an underestimate of the potential clinical situationdue to the rapid growth rate of this cancer. Previous experimentsindicate that the amelanotic melanoma increases in diameter by1 mm/day when grown on the iris (32). It would be interesting todetermine HPD concentrations in a tumor with a slower (andtherefore clinically relevant) growth rate. Fluorescence microscopy of malignant tissue showed that the highest amount ofHPD (at all time points examined) was in what appeared to benecrotic areas of the tumor. The presence of phagocytic macrophages in necrotic areas of the tumor may play a role in theobserved high level of HPD fluorescence in this area. However,histopathological examination of PDT-treated tumors show rapid

and severe vascular damage, which suggests that significantlevels of HPD are present in these well-oxygenated areas of

tumors (1, 32).Our study also demonstrated that the iris and choroid-retina

accumulate significant levels of HPD while the sclera retainedlow amounts of porphyrin. HPD was not detected in avascularstructures such as the cornea and lens. HPD was rapidly takenup in the iris and was also cleared from this structure. Thepharmacokinetics of HPD in the choroid-retina shows both a

rapid uptake and a partial clearance following systemic injection.However, HPD levels in the choroid-retina remained high even

out to 48 h. Fluorescence microscopy data indicate that HPDconcentrates first in the choroid microvasculature, is taken upby the retinal pigment epithelium, and then localizes within theouter segments of the retina. The retina of the rabbit eye is notvascularized (except for the medullary ray), and the filteringproperties of the vitreous are poor. Therefore, HPD may not beeasily cleared from the retina. Instead, HPD remains within theneurosensory outer segments at concentrations equal to orhigher than that observed in tumor tissue. The effects of avascularized retina, as found in humans, on HPD localization is

not known and precludes any accurate comparison of HPDdistribution in the retina of rabbits and humans.

The low concentration of HPD found in the sclera suggeststhat transscleral light delivery may be an effective and safemethod of treating the base of ocular melanomas. Most choroidaland ciliary body melanomas are pigmented and thicker than 5mm (18). A PDT treatment which combines transpupillary andtransscleral light delivery might provide a more effective procedure for complete tumor eradication. Additional studies areneeded in this area, but it would appear from our HPD distributionexperiments that photodynamic injury to the sclera would beminimal.

The absence of HPD in the cornea and lens correlates withthe lack of observable acute or chronic damage to these structures following HPD PDT. An earlier study demonstrated thatacute ocular damage in the form of well-circumscribed areas ofretinal edema, detachment, and necrosis could be induced byHPD PDT (31). Our current study, which had a follow-up period

which ranged from 9 to 30 months, demonstrated that the retinaldamage was localized and not progressive. The ability to accurately localize the beam of light used in treating ocular tumorsshould minimize the amount of surrounding retina which is exposed and therefore the amount of normal retina at risk forpermanent damage. The facts that HPD was not detected in thelens of any experimental eyes and that lens opacities were notobserved during the follow-up examination period suggest that

HPD PDT will not produce complications such as cataracts. Inagreement with these observations is the fact that we haveobserved patients treated with ocular HPD PDT for up to 24months without evidence of any corneal or lens opacities."

However, photosensitizers such as chlorpromazine, promazine,and hematoporphyrin can induce in vitro photopolymerizationand histidine destruction in calf lens protein (38). In addition, 8-methoxypsoralen (which is used in phototherapy of psoriasis)can induce cataracts (39). Therefore, potential problems relatedto lens damage induced by HPD PDT should not be ruled out.

REFERENCES

1. Dougherty. T. J. Photodynamic therapy (PDT) of malignant tumors. CRC Crit.Rev., pp. 83-116. 1984.

2. Dougherty, T. J., Weishaupt, K. R., and Boyle, D. G. Photoradiation therapyof malignant tumors. In: V. DeVita, S. Hellman, and S. Rosenberg (eds.),Principles and Practices of Oncology, pp. 1836-1844. Philadelphia: J. B.Lippincott Co., 1982.

3. Gomer, C. J., and Dougherty, T. J. Determination of 3H and 14Chematopor

phyrin derivative distribution in malignant and normal tissue. Cancer Res., 39:146-151, 1979.

4. Weishaupt, K. R., Gomer, C. J., and Dougherty, T. J. Identification of singleoxygen as the cytotoxic agent in photoinactivation of a murine tumor. CancerRes.. 36: 2326-2339, 1976.

5. Wan, S., Parrish, J. A., Anderson, R. R., and Madden, M. Transmittance ofnonionizing radiation in human tissues. Photochem. Photobiol., 34: 679-681.1981.

6. Benson, R. C., Kinsey, J. H., Cortese. D. A., Farrow, G. M., and Utz, D. C.Treatment of transitional cell carcinoma of the bladder with hematoporphyrinderivative phototherapy. J. Urol., 730: 1090-1095, 1983.

7. Bruce. R. A. Photoradiation for choroidal malignant melanoma. In: A. Andreoniand R. Cubeddu (eds.). Porphyrins in Tumor Phototherapy, pp. 455-461. NewYork: Plenum Publishing Corp.. 1984.

8. Cortese, D. A., and Kinsey, J. H. Hematoporphyrin derivative phototherapy forlocal treatment of cancer of the tracheobronchial tree. Ann. Otol. Rhinol.Laryngol., 97: 652-655, 1982.

9. Dahlman, A.. Wile, A. G., Bums, R. G., Mason, E. R., Johnson, F. M., andBerns. M. W. Laser photoradiation therapy of cancer. Cancer Res., 43: 430-434, 1983.

4A. L. Murphree, unpublished data.


3721



HPD UPTAKE AND PHOTOSENSITI2ATION IN THE EYE

10. Dougherty, T. J., Lawrence, G., Kaufman, J. H., Boyle, D. G., Weishaupt, K.R., and Goldfarb, A. Photoradiation in the treatment of recurrent breastcarcinoma. J. Nati. Cancer Inst., 62: 231-237, 1979.

11. Dougherty, T. J., Kaufman,J. E., Goldfarb, A., Weishaupt, K. R.. Boyle, D. G.,and Mittleman, A. Photoradiation therapy for the treatment of malignanttumors. Cancer Res., 8: 2628-2635,1978.

12. Hayata, Y., Kato, H., Konaka, C., Ono, J., and Takizawa, N. Hematoporphyrinderivative and laser photoradiation in the treatment of lung cancer. Chest, 87:269-277,1982.

13. Laws, E. R., Cortese, D. A., Kinset, J. H., Eagan, R. T., and Anderson, R. E.Photoradiation therapy in the treatment of malignant brain tumors, a phase I(feasibility)study. Neurosurgery, 9: 672-677, 1981.

14. Tse, D. T., Dutton, J. J., Weingesit, T. A., Hermsen,V. M., and Kersten, R, C.Hematoporphyrin photoradiation therapy for intraocular and orbital malignantmelanoma.Arch. Ophthalmol., 702: 833-838,1984.

15. Girotti, W. W. Mechanismsof photosensitization. Photochem. Photobiol., 38:745-751,1983.

16. Kessel, D. Hematoporphyrin and HPD: photophysics, photochemistry andphototherapy. Photochem. Photobiol., 39: 851-860,1984.

17. Lewis, R. A., Tse. D. T., Phelps, C. D., and Weingeist, T. A. Neovascularglaucomaafter photoradiationtherapy for uvealmelanoma.Arch. Ophthalmol.,102:839-842,1984.

18. Brady, L. W.. Shields, J. A., Ausburger, J. J., and Day, J. L. Malignantintraocular tumors. Cancer,49: 578-585. 1982.

19. Seigel, D., Myers. M., Ferris, F., Ill, and Steinhorn, S. C. Survival rates afterenucleation of eyes with malignant melanoma. Am. J. Ophthalmol., 87: 761-765, 1979.

20. Shields,J. A. Diagnosisand Managementof IntraocularTumors. St. Louis: C.V. Mosby Company, 1983.

21. Shields, J. A., and Ausburger, J. J. Current approaches to the diagnosis andmanagementof retinoblastoma. Surv. Ophthalmol.,25: 347-372,1981.

22. Abramson, D. H., Jereb, B., and Ellsworth, R. M. External beam radiation forretinoblastoma. Bull. NY Acad. Med., 57: 787-803, 1981.

23. Mac Faul, P. A. Local radiotherapy in the treatment of malignant melanomaofthe choroid. Trans. Ophthalmol. Soc. UK, 97: 421-426, 1977.

24. Char. D. H., Lonn, L. I., and Margoiis, L. W. Complications of cobalt plaquetherapy of choroidal melanoma.Am. J. Ophthalmol.,84: 536-541, 1977.

25. Char, D. H., and Castro, J. R. Helium ion therapy for choroidal melanoma.Arch. Ophthalmol., 700: 935-938, 1982.

26. Gragondas, E. S., Goitein, M., Verhey, L., Munzenreider,J., Urie, M., Suit, H.D., and Koehler. A. Proton beam irradiationof uveal melanomas,results of 5Viyear study. Arch. Ophthalmol., 700. 928-934, 1982.

27. Cunningham, R. D., and Henderson,J. W. Experimental evaluationof hema-toporphyrin in the detection and management of intraocular tumors. Am. J.Ophthalmol., 67: 36-44, 1966.

28. Krohn, D. L., Jacobs. R., and Morris, D. A. Diagnosis of model choroidalmalignant melanoma by hematoporphyrin derivative fluorescence in rabbits.Invest. Ophthalmol.. 73: 244-255, 1974.

29. Benedict, W. F., Lingua, R. W., Doiron, D. R., Dawson, J. A., and Murphree,A. L. Tumor regression of human retinoblastoma in the nude mouse modelfollowing photoradiationtherapy: a preliminaryreport. Med. Pediatr.Oncol. 8'387-401.1980.

30. Gomer, C. J., Rucker, N., Mark, C., Benedict, W. F., and Murphree, A. L.Tissue distribution of 3H-hematoprophyrinderivative in athymic nude miceheterotransplantedwith humanretinoblastoma.Invest.Ophthalmol.VisualSci.,22: 118-120,1982.

31. Gomer, C. J., Doiron, D. R., Jester, J. V., Szirth, B. C., and Murphree, A. LHematoporphyrin derivative photoradiation therapy for the treatment of intraocular tumors: examinationof acute normalocular tissue toxicity. CancerRes.,43:721-727, 1983.

32. Gomer, C. J., Doiron, D. R., White, L., Jester, J. V., Dunn, S., Szirth, B. C.,Razum, N. J., and Murphree, A. L. Hematoporphyrinderivative photoradiationinduceddamageto normaland tumor tissue of the pigmentedrabbit eye. Curr.EyeRes., 3. 229-237, 1984.

33. Liu, L. H. S., and Chuo, N. Hematoporphyrin phototherapy for experimentalintraocular malignantmelanoma.Arch. Ophthalmol., 707: 901-903,1983.

34. Sery, T. W., and Dougherty, T. J. Photoradiation therapy of ocular malignantmelanomaconditioned by hematoporphyrinderivative.Curr. Eye Res.,3: 519-528, 1984.

35. Gomer, C. J., Doiron, D. R., Rucker. N., Razum, N. J., and Fountain, S. W.Action spectrum (620-640 nm) for hematoporphyrin derivative induced cellkilling. Photochem.Photobiol., 39: 365-368,1984.

36. El-Far, M. A., and Pimstone, N. R. Tumor localizationof uroporphyrin isomersI and III and their correlation to albumin and serum protein binding. CellBiochem. Fund., 7. 156-160, 1983.

37. Gomer, C. J., and Little, F. H. Documentation of radioactive contaminants intritium labeled hematoporphyrin derivative. In: D. R. Doiron and C. J. Gomer(eds.), Porphyrin Localization and Treatment of Tumors, pp. 591-600. NewYork: Alan R. Liss, 1984.

38. Roberts, J. E. The photodynamic effect of chlorpromazine, promazine andhematoporphyrin on lens protein. Invest. Ophthalmol. Visual Sci., 25: 746-750, 1984.

39. Lerman, S. Ocular phototoxicity and psoralen plus ultraviolet radiation (320-400 nm) therapy: an experimental and clinicalevaluation.J. Nati. Cancer Inst.,69: 287-298, 1982.

Fig. 1. Intraocular tumor 6 h following HPD (10 mg/kg) administration.A, phase-contrast micrograph of tumor depicting an irregular area of fragmented tumor cells(arrow) suggestive of necrosis. At open arrow, a small vessel is seen surrounded by intact tumor cells. 8, fluorescence micrograph of same area showing HPDfluorescenceof fragmented cells (arrow). No fluorescencewas observed in or around vessel (openarrow), x 250.

Fig. 2. Ciliary process 30 min following HPD administration. A, phase-contrast micrograph of ciliary process which is lined by a pigmented epithelial cell layer(arrowhead).Small arrow, area which fluoresces following excitation of HPD.B, fluorescencemicrographof ciliary epitheliumas well as within individualcells (capillaries).x250.

Fig.3. Choroid-retina 30 min following HPD administration. A, phase contrast of choroid-retinal depicting choroid (aslerisk), retinal pigment epithelium (arrow), andoverlying retina. B, fluorescence micrograph demonstrating HPD fluorescence localized to small choroidal vessels (smallarrow) and the retinal pigment epithelium(largearrow), x 250.

Fig.4. Choroid-retina24 h following HPD administration.A, phase-contrast micrographof choroid-retinadepicting choroid (asterisk),retinal pigment epithelium(curvedarrow), and the overlying neurosensory retinal outer segments (openarrow). B, fluorescencemicrograph of choroid-retinademonstrating HPD fluorescenceof the retinalpigment epithelium and neurosensory retinal outer segments, x 1000.


3722




3723




3724




Fig. 5. Clinical appearanceof Rabbit Eye 5 prior to treatment and at various time intervals following PDT (10 mg HPD/kg. 90 J/sq cm). A. fundus photograph takenprior to treatment, ÃŸ,fundus photograph taken 2 days posttreatment. Retinal detachment and folds are seen within the treatment field. C, fundus photograph taken 7days posttreatment. 0, 6 months posttreatment. E, 21 months posttreatment. Retinal vasculature within the medullary ray appears unchanged from pretreatmentobservations. The area of retinal damage has remainedconstant throughout the observation period.


3725



1985;45:3718-3725. Cancer Res Charles J. Gomer, James V. Jester, Nicholas J. Razum, et al. Damage in Rabbit Ocular TissueHematoporphyrin Derivative Distribution and Long-Term Photodynamic Therapy of Intraocular Tumors Examination of

Updated version

http://cancerres.aacrjournals.org/content/45/8/3718

Access the most recent version of this article at:

E-mail alerts related to this article or journal.Sign up to receive free email-alerts

Subscriptions

Reprints and

[email protected] at

To order reprints of this article or to subscribe to the journal, contact the AACR Publications

Permissions

Rightslink site. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC)

.http://cancerres.aacrjournals.org/content/45/8/3718To request permission to re-use all or part of this article, use this link



http://cancerres.aacrjournals.org/cgi/alerts

mailto:[email protected]



photodynamic therapy of intraocular tumors examination of...

Documents