Biochemical Changes Induced in Hairy-Cell Leukemia Following Treatment with the Adenosine Deaminase Inhibitor 2'-Deoxycoformycin'

(CANCER RESEARCH 46. 2179-2184, April 19861 Biochemical Changes Induced in Hairy-Cell Leukemia following Treatment with the Adenosine Deaminase Inhibitor 2'-Deoxycoformycin'

James B. Johnston,2 Asher Begleiter, Linda Pugh, Marsha K. Leith, John A. Wilkins, Douglas J. Cavers, and Lyonel G. Israels Manitoba Institute ofCell Biology, Manitoba Cancer Treatment and Research Foundation fJ. B. J., A. B., L. P., M. K. L., L. G. I.J, and Departments oflnternal Medicine If. B.J.,A.B.,L G.l.J.Immunology(J.A. W.J,andPathologyID.J. C.],UniversityofManitoba,Winnipeg,Manitoba,CanadaR3EOV9

ABSTRACT counts (3, 13, 18). In addition, this regimen is highly effective in HCL and produces complete and durable remissions in some The adenosine deaminase inhibitor 2'-deoxycoformycin and interferon patients (14, 16, 17). It is generally accepted that the lympho are highlyeffectivein the treatment of hairy-cellleukemia.In this study, cytolytic activity of dCF is mediated through the intracellular a patient with type 2 hairy-cell leukemia was treated with one cycle of 2'-deoxycoformycin (4 mg/m2, i.v. weekly for 3 weeks), which was accumulation ofdeoxyadenosine and adenosine following ADA repeated at 9 wk. No toxicity was observed,and the hairy cell count fell inhibition. Deoxyadenosine may be converted in lymphoid cells from72,000/mm3to5,000/mm3in3 mo,witha concomitant50%decrease to dATP, and this has been observed in ALL (3â€”6,8, 10), in the spleen size. The erythrocyte deoxyadenosine triphosphate content SÃ©zarysyndrome (19), CLL (20), and HCL (14) following dCF increased to 13.6 pmol/I0' cells followingthe initial three weeklytreat therapy. Furthermore, both inherent (21) and acquired (10) ments, but there was no decrease in the adenosinetriphosphate poolsize resistance of ALL to dCF has been attributed to an inability of and noevidenceof hemolysis.The hairy celladenosinedeaminaseactivity the leukemic cells to generate dATP from deoxyadenosine. was inhibited by >95% 24 h following the first 2'-deoxycoformycin dATP inhibits ribonucleotide reductase and DNA synthesis injectionand returned to the pretreatment valueat Day8, although there (22), but this feature cannot explain the activity of dCF in the was a linear decline in peripheral hairy cell count (50%) during this slowly proliferating tumors, CLL and HCL (20). Alternatively, period. No ultrastructural changes were observed in the hairy cells following 2'-deoxycoformycin to suggest lymphocytotoxicity or cellular dATP accumulation may produce a fall in cellular ATP (8, 19, differentiation. The antitumor activity of 2'-deoxycoformycin could not 23, 24), which may explain the activity of dCF in ALL (8) and be attributed to alterations in the hairy cell deoxyadenosine triphosphate/ SÃ©zarysyndrome (19). Recent studies have demonstrated that adenosinetriphosphate levelsor to the inductionof DNA strand breaks. the cytotoxicity of deoxyadenosine and dCF to human periph Additionally, the plasma levels of interferon did not change during eral lymphocytes in vitro may be related to the induction of therapy, making it unlikely that 2'-deoxycoformycinexerts its activity DNA strand breaks (25, 26), which may occur as a result of by inducingendogenousinterferon synthesis. dATP accumulation (26). Although DNA fragmentation inhib its RNA synthesis (26, 27), the lethal event appears to be the INTRODUCTION depletion of cellular NAD, which is utilized in the production of poly(ADP-ribose) required for DNA repair (26). Apart from ADA3 is responsible for the deamination of deoxyadenosine the effects on lymphocyte dATP levels, deoxyadenosine and and adenosine to deoxyinosine and inosine, respectively, and adenosine may produce an increase in the cellular levels of 5- congenital deficiency ofthis enzyme is associated with profound adenosylhomocysteine and thus induce lymphocytolysis or cel lymphopenia and severe combined immunodeficiency (primar lular differentiation by inhibiting S-adenosylmethionine-medi ily T-cell) (I, 2). This observation provided the rationale for the ated methylation (2, 10, 11, 28). In the present study we have development of ADA inhibitors for use in lymphoid cancers or evaluated the mechanism of action of dCF in a patient with as immunosuppressive agents. dCF, which is the most potent type 2 HCL by correlating the clinical response with effects of ofthese agents, having a K1of2.S x I0_12 M (1, 2), has shown therapy on the HC ADA activity, dATP/ATP pools, and the activity in T-cell ALL (3â€”11), although considerable toxicity frequency of DNA SSB. In addition, as HCL is also highly was observed, as large cumulative dosages of dCF were required responsive to interferon (29, 30), we have examined the possi to ablate the high blast cell ADA activity (3, 4, 7). Using lower bility that dCF may exert its antitumor effect in this cancer doses of dCF, partial and complete remissions have been oh through the stimulation of endogenous interferon synthesis. tamed in mycosis fungoides, with tolerable toxicity (4, 12, 13). In contrast to the T-cell cancers, the lymphocytes in CLL and MATERIALS AND METHODS HCL have low levels of ADA activity (3, 14, 15), and this activity is completely inhibited by dCF (4 mg/m2) i.v. (3, 14), Materials. RPMI Medium 1640 and fetal-bovine serum were oh which can be repeated at weekly intervals with nausea/vomiting tamed from Gibco Laboratories, Grand Island, NY. Ficoll 400 was and lethargy being the major toxicities (3, 13, 14, 16â€”18). purchased from Pharmacia Fine Chemicals, Inc., Dorval, Quebec, and Weekly dCF may produce a rapid fall in the peripheral lym sodium diatrizoate was from Sterling Organics, New York, NY. Calf phocyte count in refractory CLL, and approximately 50% of intestinal ADA, nucleosides, and nucleotides were purchased from pancytopenic patients achieve improvements in the blood Sigma Chemial Co., St. Louis, MO. Hoescht 33258 was supplied by Calbiochem-BehringCorp., Milwaukee,WI; polycarbonate filters were Received 10/2/85; accepted I2/30/85. from Nucleopore Corp., Pleasanton, CA, and proteinase K was from The costs of publication of this article were defrayed in part by the payment E. Merck, Darmstadt, Germany. of page charges. This article must therefore be hereby marked advertisement in Patient. A 65-yr-old male presented with uncomfortable splenomeg accordance with 18 U.S.C. Section 1734 solely to indicate this fact. @ Supported by a grant from the National Cancer Institute, Canada. aly and a 6-mo history of early satiety with a weight loss of 30 lb. 2To whom requests for reprints should be addressed. at Manitoba Cancer Physical examination revealed marked splenomegaly with the spleen Treatment and Research Foundation, 100 Olivia Street, Winnipeg, Manitoba tip being 17 cm below the left costal margin, but there was no hepato R3E 0V9. Canada. megaly or lymphadenopathy. The hemoglobin was 12.8 g/dl; platelet 3 The abbreviations used are: ADA, adenosine deaminase; dCF, 2'-deoxyco formycin: PBS. phosphate-buffered saline (0.8 m@iKH2PO4:0.154 MNaCI); ALL, count, 160,000/mm3; WBC, 75,000/mm3; and reticulocyte count, acute lymphoblastic leukemia; CLL, chronic lymphocytic leukemia: HCL, hairy 125,000/mm3.The WBC differential showed 4% neutrophils and 96% cell leukemia: HC. hairy cell: SSB. single strand break(s). lymphoid cells with >90% having HC morphology. The HC had 2179

2'-DEOXYCOFORMYCIN AND HAIRY CELL LEUKEMIA

centrally placed nuclei and prominent nucleoli, with clumping of the during this time it was stored at 4C. The total volume in 24 h was chromatin at the periphery of the nuclei and numerous filamentous measured, and an aliquot was prepared for deoxyadenosineanalysis, as projections (Fig. 1). Cytochemical staining showed that these cells described below. strained strongly positive for tartrate-resistant acid-phosphatase. The ADAAssay.The mononuclear cell ADA activitywas measured using bone marrow aspirate was easily obtained, and 30% of the nucleated the method ofHartwick et a!. (32) as we have previouslydescribed(14). cells were HC. A bone marrow biopsy was normocellular and demon Cells were reconstituted in PBS titrated to pH 6.8, freeze-thawed 3 strated a diffuse infiltration with HC. Immunological typing of the times, and centrifuged at 15,000 x g for 15 mm. An aliquot of peripheral mononuclear cells showed a marked increase in the number supernatant was incubated at 37'C for 30 or 60 mm with an equal of cells with B-cell surface markers and a slight increase in the number volumeof i0@ Madenosine in PBS (pH 6.8). The reaction was stopped with T-cell markers. The absolute numbers were: cells with surface with an equal volume of ice-cold 10% trichloroacetic acid, the mixture immunoglobulins, 53,655/mm3 (normal, 47â€”638);cellswith Bl anti was filtered through 0.5-tim Fluoropore filters (Millipore), and it was gen, 52,000/mm3 (normal, 130â€”342);cellsforming sheep erythrocyte neutralized with 2 volumes of 0.5 trioctylamine in trifluorotrichloro rosettes, 7,580/mm3 (normal, 285â€”2,148); and cells with T3 antigen, ethane (33). The conversion of adenosine to inosine and hypoxanthine 3,500/mm3 (normal, 1,008â€”1,914). Serum immunoelectrophoresis was measured by reversed-phase high-pressure liquid chromatography, demonstrated a normal IgG level at 9.21 g/liter (normal, 6.6â€”16.47); using BrownleeRP-l8 Spheri-5 3-cm and 10-cmcolumns connected in normal IgA, 1.10 g/liter (normal, 0.86â€”3.70),anddepressed 1gM,0.2 tandem. The mobile phase was 0.01 M KH2PO4 (pH 3.8):15% (v/v) g/liter (normal, 0.61â€”3.09).There was no serum paraprotein and no methanol, and the flowrate was I.5 ml/min. Peak areas were measured Bence Jones proteinuria. Liver and renal function tests were normal. by a Hewlett Packard Model 3390 A integrator and compared with Although the peripheral HC count and spleen size did not change over standards. The ADA activity was expressed as nmol of adenosine per a 6-wk period, therapy was initiated with dCF because of persistent h/b' cells (32). discomfort from splenomegaly.2'-Deoxycoformycin wasobtained from ATP and dATP Determinations.Nucleotides wereextracted from 200 the Investigational Drug Branch, National Cancer Institute, Bethesda, @lofpacked erythrocytes, or iO@peripheral mononuclear cells, with MD, and written informed consent was obtained from the patient prior 200 Mlof ice-cold 0.4 M perchloric acid and neutralized with 2 volumes to therapy. 2'-Deoxycoformycin was reconstituted in 50 ml of Ringer's of 0.5 Mtrioctylamine in trifluorotrichloroethane (33). The dATP was lactate, containing NaHCO3 (44.6 meoJliter), and was infused over 30 measured by the DNA polymerase assay of Hunting and Henderson mm. The patient received a cycle of dCF (4 mg/m2, i.v. weekly for 3 (34) and the ATP by high-pressure liquid chromatography using a times) which was repeated at 9 wk. Partisal 10 SAX anion exchange column (Whatman) (19). Isolation of Urine and Blood Components. Peripheral blood was Adenosine and Deoxyadenosine Measurements. Aliquots of urine and collected at 4C in heparinized tubes and spun at 400 x g for 10 mm, plasma were filtered through Amicon CF-25 ultrafiltration membrane and the plasma was removed and stored at â€”20'C.Thebuffy coat was cones (M@ 25,000 exclusion limit) at 750 x g for 25 mm, and the aspirated, and an aliquot of packed erythrocytes was removed for d.ATP deoxyadenosine and adenosine in the filtrate were measured by re analysis. The mononuclear cells were isolated from the huffy coat using versed-phase high-pressure liquid chromatography, as previously de a Ficoll gradient (31), followed by one wash with ice-cold PBS (pH 7.4) scribed (19). The identity of the deoxyadenosine and adenosine peaks and a second wash with ice-cold 10 mM Tris:l40 mM NH4C1 to lyse was confirmed by the disappearance ofthe peaks followingthe addition residual erythrocytes (19). Urine was collected over 24-h periods, and of excess calf intestinal ADA.

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2'-DEOXYCOFORMYCIN AND HAIRY CELL LEUKEMIA DNA SSB. DNA SSB were measured in the mononuclear cells using a fluorescent modification (35) of the alkaline elution assay described by Kohn et al. (36, 37). In these studies, sodium EDTA (pH 12.2) was used instead of tetrapropylammonium hydroxide to elute the DNA because of the high fluorescent blanks resulting from the latter base. The DNA from approximately 2 x 106 cells was eluted from 0.8-him polycarbonate filters following 1-h treatment of the lysed cells on the filter with proteinase K (0.5 mg/ml). The DNA in the eluted fractions 11 was determined by neutralizing each sample with N hydrochloric acid, adding a I-ml aliquot of the neutralized sample to 2 ml of a solution of 0.45 MMHoechst 33258 dye in 25 mM sodium phosphate buffer (pH depression excretion of deoxyadenosine were 1.5- to 2-fold greater follow in granulocyte or platelet counts or hemoglobin level. There ing the second and third dCF treatments than following the was no hemolysis during therapy, as evidenced by a change in first treatment. the reticulocyte count or serum haptoglobin, bilirubin, or lactate Cellular Nucleotide Levels. The influence of dCF on the dehydrogenase concentrations. erythrocyte and HC dATP and ATP pools is shown in Fig. 3. Hairy-Cell Morphology. No change was observed in the HC Following the first dCF treatment, the erythrocyte dATP con morphology, and there was no increased number of smudge centration gradually rose to 8 pmol/106 cells at Day 4 and cells, when the peripheral blood was examined by light micros increased to a level of 13.6 pmol/10@ cells following the two copy each day following the first dCF injection and 24 h subsequent injections. However, this change did not influence following the second and third weekly injections. In addition, the erythrocyte ATP pool size. There was no alteration in the when examined by electron microscopy 24 and 72 h following HCcontentofdATPor ATPduringdCFtherapy.Thepretreat the first dCF injection, no ultrastructural abnormalities were ment HC dATP concentration was 4 pmol/10@ cells, and the apparent in the HC to suggest cell death or differentiation (Fig. ATP level was 3000 pmol/10@ cells (peripheral mononuclear I). cell ATP concentrations in the two controls were 710 and 740 Hairy-Cell ADA Activity. The pretreatment HC ADA activity pmol/10@ cells). of 14 nmol of adenosine per h per 106 cells was less than the DNA Single Strand Breaks. DNA SSB in HC cells were control peripheral mononuclear cell ADA activities (range, 70â€” measured by alkaline elution prior to therapy, 24 and 72 h 200 nmol of adenosine per h per 10@cellsin eight controls). following the first dCF treatment and 24 h following the second The activity was < 5% of the pretreatment level 24 h following treatment. As shown in Fig. 4, no SSB were detected 24 h dCF (4 mg/m2), but increased to 60% and 100% ofthe pretreat following the first treatment, and insignificant numbers (<50 2181

0 U 0

0 aE

0 0 > 0 -j 0 V 0 0 U z Vokimeekited (ni) S Fig. 5. The alkaline elution kinetics of the peripheral mononuclear cells from the patient with HCL and two controls (A and B). Cells were incubated for 24 h 0 with dCF (50 tiM) and 1, 5, or 50 zM deoxyadenosine (dAdo) and subjected to C) alkaline elution as described in â€œMaterialsandMethods.â€•

Table 1 Frequency ofDNA SSB and dATP accumulationfoiowing exposure of mononuclear cells to dCF and deoxyadenosine in vitro DAYS Peripheral mononuclear cells were isolated from the patient, and from two Fig. 3. The hairy-cell and erythrocyte dATP and ATP levels following the controls, using a Ficoll-Paque gradient and were incubated (10' cells/ml) for 24 initial three weekly treatments with dCF (4 mg/m2). h with dCF (50 @zM)and1, 5, or 50 @Mdeoxyadenosine. The frequency of DNA SSB and the cellular dATP were measured as described in â€œMaterialsand Methods.â€• Cellular SSB (rad equivalents) and dATP content (pmol/lO' cells)

1@zM5@M5OILMdeoxyadenosinedeoxyadenosinedeoxyadenosine+dCF+dCF+dCFSubjectdATP

SSBPatient2.5Â±2.5a SSBCIATP SSBdATP 5Â±5ControlA8.1 23Â± 10â€•24.0Â±3.0 24Â±22193Â±32 278Â±62ControlB2.0Â±0.4Â±0.8 58Â±3013.0Â± 1.7 274Â±9286Â±4 117Â±247.2Â±0.9 200Â±8550Â±4 379Â±110

V a Mean Â±SE of three duplicate experiments. 0 C b Mean Â±SE of two to four experiments. S S

.( three concentrations of deoxyadenosine are shown in Fig. 5 and Table 1. Neither deoxyadenosine (50 @m)nor dCF (50 @m) alone induced DNA SSB or dATP accumulation in the HC or U S control mononuclear cells. In the control cells, the frequency U. of DNA SSB and the cellular concentration of dATP were dependent on the extracellular concentration of deoxyadeno sine. In contrast, an insignificant number of DNA SSB was observed at all deoxyadenosine levels in HC. There was no correlation between the cellular concentration of dATP and the frequency of DNA SSB as Control A cells accumulated more dATP than Control B cells, whereas Control A cells had a lower frequency of DNA SSB, and the HC accumulated 2- to 4-fold more dATP than the control cells with S or SO @mof deoxyadenosine. Interferon Levels. Plasma interferon levels were measured 0 12 24 36 each day following the first dCF injection and 24 h following Volume eluted (ml) the second and third weekly treatments. However, no increase Fig. 4. Alkaline elution kinetics of the peripheral hairy cells before therapy and I and 3 days following dCF (4 mg/m2). Cells were subjected to alkaline in the interferon levels was observed at these times. elution as describedin â€œMaterialsandMethods.â€•Included are the elution profiles produced by 400 rad. DISCUSSION rad equivalents) were detected 72 h following the first treat The patient described in this paper had type 2 HCL, which ment. No SSB were observed 24 h following the second treat differs from typical HCL by having a high peripheral HC count, ment. an easily aspirable bone marrow, relatively normal granulocyte In Vitro Incdbations. The cellular dATP accumulation and or platelet counts and hemoglobin level, and the absence of the frequency of DNA SSB following 24-h incubation of HC, monocytopenia (39â€”41).Asobserved with typical HCL (14, 16, or control peripheral mononuclear cells, with dCF (50 tim) and 17), this HCL variant is highly responsive to dCF with the 2182

Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1986 American Association for Cancer Research. 2'-DEOXYCOFORMYCIN AND HAIRY CELL LEUKEMIA peripheral HC count falling by 40â€”50%1 wk following each homocysteine concentration, with a subsequent impairment of injection of dCF (4 mg/m2). The rise in plasma and urinary RNA methylation. This feature has thus been proposed to deoxyadenosine, which reflects cell death (2, 7), was greater explain the lymphocytolysis observed following dCF in ALL following the second and third treatments compared to the first and also the occasionally observed phenotypic conversion of treatment, suggesting enhanced cell death with cumulative ALL to acute myeloidleukemia(11, 43). However,in the treatments. The plasma adenosine level also increased after present patient this mechanism appears unlikely, as only a each treatment, but due to its slower rate of urinary clearance minimal and transient elevation in plasma deoxyadenosine and (7), it fell more gradually than the deoxyadenosine level. As in adenosine was observed following dCF. (b) dCF may induce typical HCL (14), the HC in this patient had lower ADA activity lymphocytolysis by mechanisms apart from ADA inhibition as than that of normal peripheral mononuclear cells, and the ADA it has been shown that dCF is phosphorylated in human T activity was inhibited by >95% 24 h following dCF (4 mg/m2). lymphoblastoid cells in vitro and may be incorporated into However, the enzyme activity rapidly returned, despite a linear DNA (44). It is unlikely that the rapid lymphocytolysis in HCL fall in the HC count for 7 days following dCF (Fig. 2), indicating following dCF is related to the incorporation ofdCF into DNA, that the biochemical change required for cell lethality occurred as the rate of DNA synthesis in this cancer is very slow. shortly after dCF administration. The most widely accepted However, the phosphorylated derivatives of dCF may, by un biochemical perturbation to account for dCF activity is the known mechanisms, be responsible for cell death. (c) dCF may accumulation ofdATP in lymphoid cells (3â€”6,8,10, 19), which not be directly cytocidal to the HC but function, like interferon, may cause cell death through the depletion of cellular ATP (8, as a biological modifier. The present study confirms that dCF 19, 23, 24). One patient with leukemic phase HCL has been is not an inducer of interferon synthesis, but it perhaps has a described in whom the decline in HC count was preceded by similar effect as interferon on specific lymphoid cell popula the accumulation of >200 pmol dATP per 106 HC (14). This tions, which control the proliferation of HC. Recent studies patient had atypical HCL in having an 1gM paraprotein and a have demonstrated that the activity of interferon in HCL may nodular marrow infiltrate (42). In the present case the patient's be related to an increased production of natural killer cells (30), HC had a greater capacity than normal peripheral mononuclear and a similar mechanism may also explain the activity of dCF. cells to accumulate dATP when exposed to dCF and deoxy The major toxicity with weekly dCF (4 mg/m2) is nausea/ adenosine in vitro, but no alteration in the peripheral HC dATP vomiting or lethargy, and the incidence and severity of these occurred following therapy (Fig. 3). These findings indicate that toxicities increase with the number of treatments, suggesting a the HCL variants have differing capacities to accumulate dATP cumulative drug effect (14, 16, 18), which may be related to the following dCF therapy, but this feature is not essential for intracerebral accumulation of deoxyadenosine and/or adeno lymphocytolysis. In addition, the HC ATP level did not fall sine (4). Thus, the present patient received only three weekly during therapy and was initially 4-fold higher than the level in treatments, and this was repeated after an interval of 7 wk to control peripheral mononuclear cells, demonstrating that the allow recovery from the initial therapy. As with other HCL type 2 HCL has a high metabolic capacity. This fact was also patients, no toxicity was observed with this treatment schedul evidenced by the presence of prominent nucleoli (Fig. 1) and ing (14). Weekly dCF in HCL may also produce a fall in the gradual increase in HC count between treatment cycles. It hemoglobin that has been attributed to the depression of mar has recently been proposed that the lymphocytolytic activity of row erythropoiesis, which is severely compromised at this time dCF is mediated through the induction of SSB (25, 26), and due to tumor infiltration (14, 16). However, dCF-induced he this postulate was based on the observation that the cytotoxicity molysis was not excluded, and this has been observed with high of deoxyadenosine and dCF to peripheral lymphocytes was doses of dCF in SÃ©zarysyndrome and related to erythrocyte closely related to the induction of DNA strand breaks. In the ATP depletionsubsequenttodATPaccumulation(19). present study peripheral mononuclear cells developed a signif In the present patient, the erythrocyte dATP increased to icant number of DNA SSB when incubated for 24 h with 1 @M 13.6 pmol/10@ cells following the third weekly treatment, but deoxyadenosine and 50 @tMdCF, and the frequency of SSB there was no significant fall in the ATP level and no evidence increased with higher levels of deoxyadenosine and cellular of hemolysis. However, the cumulative increase in the erythro dATP accumulation (Table 1). In contrast, although the HC cyte dATP pool following each treatment (Fig. 3) suggests that accumulated 2- to 4-fold more dATP than normal mononuclear a prolonged course of weekly dCF may cause sufficient dATP cells, they did not develop SSB, suggesting that either dATP is accumulation to induce hemolysis. not responsible for SSB, or that the HC have an enhanced In summary, dCF is highly effective in type 2 HCL, but the ability to repair breaks. Regardless, DNA breakage does not antitumor activity cannot be related to effects on dATP accu appear to be responsible for the activity of dCF in HCL type 2, mulation or the induction of DNA strand breaks in the HC. as no SSB were observed in the HC following therapy (Fig. 5). Further studies are required to determine the metabolism of Other biochemical mechanisms may account for the activity dCF in HC and examine its role as a biological modifier. ofdCF in HCL type 2. (a) The accumulation of deoxyadenosine and adenosine following ADA inhibition may increase the ACKNOWLEDGMENTS cellular level of S-adenosylhomocysteine and inhibit essential S-adenosylmethionine-mediated transmethylation reactions (2, The authors wish to thank Joan Powell for the typing of this 10, 11, 28). Deoxyadenosine inactivates S-adenosylhomocy manuscript. steine hydrolase, which degrades S-adenosylhomocysteine to adenosine and homocysteine, and an increase in adenosine also REFERENCES produces S-adenosylhomocysteine accumulation, as the equilib I. Agarwal, R. P. Inhibitors of adenosine deaminase. Pharmacol. Ther. Part A rium constant for this reaction favors the catabolic direction Chemother.Toxicol.Metabl.Inhibitors, 17: 399â€”429,1982. (2). Hershfield et a!. (10) have shown that high doses of dCF in 2. Kredich, N. M., and Hershfield, M. S. 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Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1986 American Association for Cancer Research. Biochemical Changes Induced in Hairy-Cell Leukemia following Treatment with the Adenosine Deaminase Inhibitor 2 ′ -Deoxycoformycin

James B. Johnston, Asher Begleiter, Linda Pugh, et al.

Cancer Res 1986;46:2179-2184.

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