Intramitochondrial lamellar bodies in acute myeloblastic leukemia

Intramitochondrial Lamellar Bodies in Acute Myeloblastic Leukemia ROBERTO HERRERA-GOEPFERT,MD,* ROBERTO BARRIOS-DELVALLE, MD,* VICTOR SALES-CARMONA, MD,t JORGE SANTOYO, MD,t AND ELENA B. OLIVA-RAMIREZ, BSc*

Intramitochondrial lamellar bodies w e r e o b s e r v e d in three cases of acute myeloblastic leukemia. Two of the patients had M1 leukemia a n d the remaining patient M4 leukemia, by the FAB classification. In all three cases neoplastic ceils c o n t a i n e d dilated mitochondria that varied in size a n d s h a p e and c o n t a i n e d d e c r e a s e d numbers of cristae. Some mitochondria contained lamellar structures that resembled myelin figures and, occasionally, primary granules; these structures were more conspicuous in the central portion of the mitochondria. Regardless of the proliferating cell type (lymphoblasts, myeloblasts, or monoblasts), there a r e common ultrastructural changes that represent abnormal metabolic function, such as d i s o r d e r s of intramitochondrial protein synthesis. The exact meaning of these findings is not known; adequate interpretation will require further investigation of the biology o f t h e s e neoplastic processes. HUM PATHOL 17:748--753, 1986.

MATERIALSAND METHODS

From each of three patients with a clinical diagnosis of acute myeloblastic leukemia, M 1 in two and M4 in one, we obtained 5 ml of venous peripheral blood. The patients were 19, 21, and 52 years of age and had not been treated previously. All samples were collected in test tubes that contained 30 IU of heparin/ml (150 IU each). Buffy coat was prepared

Acute leukemias have been classified according to the proliferating cell type as lymphoblastic (ALL) and myeloblastic (AML) leukemias. This classification is also important because of its therapeutic and prognostic implicationst; however, in some cases, predominantly those labeled as undifferentiated leukemias, the differential diagnosis is difficult. On the basis of past experience it has been possible to identify specific disorders in cell metabolism. Several teclmiques directed to the characterization of enzymatic and surface markers have been extremely usefitl in the identification of particular neoplastic populations. ~-6 Uhrastructural analysis by the buff), coat method has contributed valuable information.7-0 The morphologic changes, some of them constant, that have been described permit the distinction between types of leukemia with relative certainty. In an uhrastructural prospective study of leukenfic cells that we recently began in our laboratory, we found intramitochondrial lamellar bodies in three cases of acute myeloblastic leukemia; in one case intranuclear bodies were also found. T h e importance of these findings, which to our knowledge have not been described previously, prompted us to make some comments as to their possible meaning. Received from tile *Unidad de Microscopia Electronica, Facuhad de Medicina, UNAM, and the tServicio de Hematologia, Hospital General de Mexico, Secretaria de Salud. Revision accepted for publication December 31, 1985. Address correspondence and reprint requests to Dr. tterreraGoepfert: Depto. de Patologia, Instituto Nacional de Cardiologia "Ignacio Chfivez,"Juan Badiano No. 1 (lntersecci6n Perif6rico Sur y Viaducto Tlalpan), Col. Secci6n XVI, Delegaci6n Tlalpan CP 14080, M6xico, DF Mexico.

748

FIGURE 1 (top]. One-micron section from a patient with acute myeloblastic leukemia, M1. There are multiple cells with moderate amounts of cytoplasm, peripherally distributed chromatin, and excenfric nucleoli, ffoluidine blue stain, x 800.] FIGURE 2 [bottom]. One-micron section from leukemic cells from a patient with acute myeloblastic leukemia, M4. Monocytoid cells are intermingled with myelobasts. [loluidine blue stain, x 800.]

ACUTE MYELOBLASTICLEUKEMIA(Herrera-Goepfert et aL)

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FIGURE 3 (top). Cells from a patient with acute myeloblastic leukemia, MI. In most cells organelles are scarce; dilated mitochondria are present. Bundles of intranuclear filaments are seen in some cells [arrows). {x 5,500.} FIGURE 4 (bottom]. High-power view of a cell from a patient with acute myeloblastic leukemia, M'I, in which enlarged ond pleomorphic mitochondria are present. There are numerous rosettes of ribosomes and dilated cisternae of rough endoplasmic reticulum. [ x 17,600.]

and fixed in 2.5 per cent glutaraldehyde in cacodylate buffer, 0.067 M, pH 7.4. Samples were then washed ill the same buffer, postfixed in 1 per cent osmium tetroxide, dehydrated in ethanol, and emb e d d e d in Epon 812. Sections cut at 1 lira were stained with toluidine blue, and specific areas were selected for the preparation of tlfin sections, which were stained with lead citrate and uranyl acetate and examined in an electron microscope. 749

RESULTS

Light Microscopic Findings In all cases neoplastic cells were regular in size, with moderate amounts of cytoplasm; occasionally, small vacuoles were present; we did not identify grannies. Nuclei were llomogeneous, with peripherally distributed and finely clumped cbromatin; conspicuous nncleoli and shallow notches were present

HUMAN PATHOLOGY

Volume 17, No. 7 [July 1986]

FIGURE 5 [top]. Cell in whlch perinuclear mitochondria reveal loss of cristae and formation of lamellar structures. Bundles of filaments are visible In intranuclear and intracytoplasmic locations, [ x t7,600.] FIGURE 6 [bottom]. High-power view of a dense lamelIar structure with an internal structure somewhat similar to that of a specific type of primary granule. [ x 55,000,]

found (fig. 3). Rosettes of ribosomes and some dilated cisternae of rongh endoplasmic reticulum were seen (fig. 4). Dense lamellar structures, some of them in a primary granule-like configuration, were observed within some mitochondria (figs. 5 and 6); occasionally, azurophil granules were present. Regular nuclei contained peripheral heterochromatin and prominent nucleoli. In one o f the patients with M1

in some cells (fig. 1). In addition, examination of some cells from the patient with M4 leukemia revealed a monocytoid pattern (fig. 2).

Electron Microscopic Findings Cells of A M L - M I had regular profiles with scarce organelles in which dilated mitochondria were 750

ACUTE MYELOB[ASTIC LEUKEMIA (Herrera-Goepfert et al.)

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tures, such as virus-like particles, have also been described within the mitochondria of leukemic cells. 12,16 In any event, there are experimental models in which tile mitochondrial damage is similar to that observed in leukemic cells.]7.]s Cumulative knowledge o f mitochondrial metabolism 19,2~ allows us to assume, on a morphologic basis, that our observations could be interpreted as disturbances in the structure of the inner membrane due to defects in intramitochondrial protein synthesis, in addition to a defect in the transportation and uptake o f certain cytoplasmic proteins; similar, although not identical, azurophil granule-like strnctures have been described in well-differentiated polymorphonuclear leukocytes.21,22 These findings are, in our opinion, an example of what is known as asynchronism and may reflect an attempt toward spontaneous maturation o f the granulocytic series. 2 3 ~ er think that it would then be a very useful morphologic marker in the diagnosis and identification of patients with myeloid leukemias that initially resemble undifferentiated leukemias. On the other hand, the presence in one of our cases o f lamellar structures with the nuclear characteristics described earlier is in agreement with previous observations of marked alterations in nuclear and mitochondrial DNA. Fibrillar structures have been observed in the nt, clei and cytoplasm of neoplastic cells of other types. 24 In leukemic cells they have been reported in an intranuclear location only in neoplastic myeloblasts and monoblasts, which, according to one study, could share a common origin. 25 As mentioned earlier, these structures could be a neoplastic marker for leukemic cells of myeloid or-

leukemia, intranuclear and intracytoplasmic filaments were arranged in bundles (fig. 5), and lamellar structures were present within the nucleus (fig. 7). In the patient with M4 leukemia there was a mixture of cells, some with the characteristics mentioned earlier and others with a monocytoid appearance, with shallow indentations, scarce pleomorphic granules, and phagocytic vacuoles (fig. 8). Within the mitochondria of both cell populations intramitochondrial lamellar structures were observed (fig. 9).

DISCUSSION Leukemic cells are blood elements that have been transformed both metabolically and morphologically. They have a capacity for widespread proliferation, explaining, at least in part, their rapid dissemination and infiltration o f many organs. Several methods, in addition to morphologic criteria, have been applied and established for the identification and differentiation of leukemias of myeloblastic and lymphoblastic origin. These methods include deterruination of enzymatic and surface markers by both light and electron microscopy, l~ Quantitative uhrastructural studies are also helpful in the correct interpretation of the cell type. ~z Furthermore, there is evidence that morphologic changes reflect abnormal cell metabolism; such changes have been related to defects in the biosynthesis of nucleic acids by modified genes located in some chromosomes. 1~ On the other hand, it has been demonstrated that mitochondrial DNA is also altered, and this change has been correlated with decreased numbers ofcristae and vacuolated mitochondria. 14'15 O t h e r interesting fea-

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HUMAN PATHOLOGY

Volume 17, No. 7 (July 1986)

FIGURE 8 [top]. Cells from a patient with acute myeIoblastic leukemia, M4; marked pleomorphism of cytoplasmic granules is seen. An intramitoohondrial granule-like structure is visible [arrow]. ( x 5,500.] FIGURE 9

[bottom]. Lamellar structures in a concentric arrangement in an intramitochondrial location. [ x 35,200.)

igin a n d could be helpfill in cases in which determination o f cell line is difficult; t h e r e f o r e , they should be carefully sought. Even then, the significance o f these structures requires the application o f m e t h o d s o t h e r than m o r p h o l o g i c studies alone, as well as further observations, to elucidate their real origin and constitution.

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