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Hemolytic Anemias

Chapter 105 | Part 4: Oncology and Hematology · Part 4 – Oncology: Hematologic Malignancies · Chapter 105


Key Clinical Points

  1. Reticulocyte count is the definitive parameter for hemolysis; both percentage (>2%) and absolute count (>150,000/μL) are typically increased.
  2. Hemolytic anemias (HAs) are classified as inherited or acquired, and intracorpuscular (membrane, enzyme, hemoglobin) or extracorpuscular (immune, mechanical, etc.).
  3. Hereditary spherocytosis (HS) prevalence is 1:2000–5000 in European ancestry; splenectomy is indicated for severe cases (Hb <8 g/dL) at age 4–6 years.
  4. G6PD deficiency is X-linked; heterozygous females show variable expression due to X-chromosome inactivation mosaicism.
  5. Pyruvate kinase (PK) deficiency is autosomal recessive; mitapivat (10 mg/kg/day) is an allosteric activator of PK.
  6. Chronic extravascular hemolysis leads to iron overload (secondary hemochromatosis) and organ damage (liver, heart).
  7. Osmotic fragility testing is the primary diagnostic for HS; EMA-binding test identifies ankyrin deficiency.
  8. Splenectomy is strictly contraindicated in stomatocytosis due to severe thromboembolic complications.
  9. Parvovirus B19 infection can cause aplastic crisis in patients with chronic hemolysis (e.g., PK deficiency).
  10. Compensated hemolysis may be asymptomatic; decompensation occurs during pregnancy, folate deficiency, or renal failure.

1. DEFINITION & OVERVIEW

Hemolytic anemias (HAs) are defined as anemias resulting from increased destruction of red cells.

Core Features: ◦ Increased peripheral blood red cell consumption with normal or elevated marrow production. ◦ Differentiation from blood loss: Hemolysis occurs within the body; blood loss involves physical removal. ◦ Classification by etiology: Inherited/acquired; intracorpuscular/extracorpuscular. ◦ Clinical presentation varies by severity and hemolysis site (intravascular/extravascular).

Definition (Harrison's 22e): 'Hemolytic anemias (HAs) are anemias resulting from increased destruction of red cells.'

1.1 General Clinical and Laboratory Features

Clinical manifestations include:

• Jaundice, splenomegaly, gallstones, and hemoglobinuria (intravascular hemolysis). • Bone marrow overactivity may cause skeletal changes in severe congenital cases.

Laboratory features:

• Elevated unconjugated bilirubin, LDH, and reticulocytes (percentage >2% or absolute >150,000/μL). • Reduced haptoglobin in intravascular hemolysis. • Macrocytic blood smear with polychromasia and nucleated red cells. • Bone marrow aspirate typically shows erythroid hyperplasia unless otherwise indicated.

Table 105-1: Classification of Hemolytic Anemias | Classification | Intracorpuscular Defects | Extracorpuscular Factors | | --- | --- | --- | | Inherited | Hemoglobinopathies, Enzymopathies, Membrane-cytoskeletal defects | Paroxysmal nocturnal hemoglobinuria (PNH), Familial (atypical) hemolytic-uremic syndrome | | Mechanical destruction | | Drugs, Infectious | | Microangiopathic | | | | Autoimmune | | |

Table 105-2: Features Common to Most Patients with a Hemolytic Disorder | Feature | General Examination | Other Physical Findings | Hemoglobin Level | MCV, MCH | Reticulocytes | Bilirubin | LDH | Haptoglobin | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Jaundice, Pallor | Spleen may be enlarged | Bossing of skull in severe congenital cases | From normal to severely reduced | Usually increased | Almost always increased (mostly unconjugated) | Increased (up to 10× normal with intravascular hemolysis) | Reduced to absent if hemolysis is at least in part intravascular |


2. EPIDEMIOLOGY

Population-specific prevalence:

Hereditary spherocytosis (HS): 1:2000–5000 in European ancestry. • Hereditary elliptocytosis (HE): 1:2000–4000 globally. • Pyruvate kinase (PK) deficiency: 1:10,000 in most populations. • G6PD deficiency: High prevalence in malaria-endemic regions (Africa, Mediterranean, Southeast Asia). • Southeast Asia ovalocytosis (SAO): Up to 5–7% in Papua New Guinea and neighboring regions.


3. ETIOLOGY & PATHOPHYSIOLOGY

Red cell components and hemolysis mechanisms:

Red cell structure: Hemoglobin, membrane-cytoskeleton complex, and metabolic machinery. • Membrane-cytoskeleton abnormalities: Cause mechanical instability and reduced deformability (HS, HE, stomatocytosis). • Enzyme deficiencies: Disrupt ATP production or oxidative defense (PK, G6PD).

3.1 Inherited Membrane-Cytoskeleton Disorders

Genetic defects in membrane-cytoskeleton proteins:

SPTA1/β-spectrin mutations: HS (dominant or recessive), HE (dominant). • ANK1/ankyrin mutations: HS (dominant; accounts for majority of cases). • SLC4A1/band 3 mutations: HS (dominant), Southeast Asia ovalocytosis (dominant; provides protection against malaria). • PIEZO1/KCNN4 mutations: Dehydrated hereditary stomatocytosis. • RHAG/ABCB6 mutations: Chronic nonspherocytic hemolysis, pseudohyperkalemia.

Table 105-3: Inherited Diseases of the Red Cell Membrane-Cytoskeleton Complex | Gene | Chromosomal Location | Protein Produced | Disease(s) with Certain Mutations (Inheritance) | Comments | | --- | --- | --- | --- | --- | | SPTA1 | 1q22-q23 | α-Spectrin | HS (recessive), HE (dominant) | Mutations account for ~65% of HE; severe forms may involve coexisting mutations. | | ANK1 | 8p11.2 | Ankyrin | HS (dominant) | May account for majority of HS cases. | | SLC4A1 | 17q21 | Band 3 (AE1) | HS (dominant), Southeast Asia ovalocytosis (dominant) | Polymorphic mutation provides malaria resistance in heterozygotes. | | PIEZO1 | 16q23-q24 | Mechanosensitive ion channel | Dehydrated hereditary stomatocytosis (dominant) | Associated with perinatal edema and pseudohyperkalemia. | | KCNN4 | 19q13.31 | KCNN4 (Gardos channel) | Dehydrated hereditary stomatocytosis (dominant) | Clinical presentation similar to that of PIEZO1 mutants. | | ABCB6 | 2q35-q36 | ABCB6 | Familial pseudohyperkalemia (dominant) | | | RHAG | 6p21.1-p11 | Rhesus-associated glycoprotein | Chronic nonspherocytic hemolysis (recessive) | Associated with total loss of all Rh antigens; specific mutations cause overhydrated stomatocytosis. |

3.2 Enzyme Abnormalities

Key enzyme defects in red cell metabolism:

Pyruvate kinase (PK) deficiency: Autosomal recessive, 1:10,000 prevalence. • G6PD deficiency: X-linked; triggers hemolysis via oxidative stress (favism, infections). • Other rare enzymes: Hexokinase (HK), phosphofructokinase (PFK), glyceraldehyde-3-phosphate dehydrogenase (GAPD).

Table 105-4: Red Cell Enzyme Abnormalities Causing Hemolysis | Enzyme (Acronym) | Gene Symbol; Chromosomal Location | Prevalence | Clinical Manifestations | Extra-Red Cell | Comments | | --- | --- | --- | --- | --- | | Hexokinase (HK) | HK1; 10q22 | Very rare | | May benefit from splenectomy; BMTc | | G6PI | GPI; 19q31.1 | Rare | NM, CNS | May benefit from splenectomy | | PFK | PFKM; 12q13 | Very rare | Myopathy; myoglobinuria | | | Aldolase | ALDOA; 16q22-24 | Very rare | Myopathy | | | TPI | TPI1; 12p13.31 | Very rare | CNS (severe), NM | | | GAPD | GAPDH; 12p13.31 | Very rare | Myopathy | | | BPGM | BPGM; 7q33 | Very rare | Erythrocytosis rather than hemolysis | | | PGK | PGK1; Xq21.1 | Very rare | CNS, NM | May benefit from splenectomy; BMTc | | PK | PKLR; 1q22 | Rare (2) | | May benefit from splenectomy; BMTc | | G6PD | G6PD; Xq28 | Common (1) | | In almost all cases, only AHA from exogenous trigger | | GSS | 20q11.22 | Very rare | CNS | | | GSR | 8p12 | Very rare | Cataracts | | | GCLC | 6p12.1 | Very rare | CNS | | | CYB5R3 | 22q13.2 | Rare | CNS | | | AK | AK1; 9q34.11 | Very rare | CNS | May benefit from splenectomy | | P5N | NTSC3A; 7p14.3 | Rare (3) | | May benefit from splenectomy |


4. CLINICAL FEATURES

Clinical manifestations vary by hemolysis type:

Compensated hemolysis: Asymptomatic with normal hemoglobin and elevated reticulocytes. • Decompensated hemolysis: Anemia, jaundice, splenomegaly, gallstones, and complications (iron overload, aplastic crisis).

Differential diagnosis considerations:

• Acute vs. chronic presentation. • Intravascular vs. extravascular hemolysis (hemoglobinuria vs. unconjugated bilirubin).

4.1 Compensated Hemolysis versus Hemolytic Anemia

Compensated state:

Normal hemoglobin with elevated reticulocytes (count >2% or absolute >150,000/μL). • No symptoms but may show mild anemia in decompensation triggers (pregnancy, folate deficiency, renal failure).


5. DIFFERENTIAL DIAGNOSIS

Key differentials include:

AIHA vs. HS: Distinguish autoimmune hemolytic anemia from hereditary spherocytosis. • G6PD vs. other enzyme defects.PNH vs. cold agglutinin disease.MAHA vs. mechanical destruction.

Table 105-8: Classification of Acquired Immune Hemolytic Anemias | Clinical Setting | Type of Antibody | | --- | --- | | | Cold (mostly IgM, 4°C–30°C) | Primary: CAD; Secondary to infection (Mycoplasma → PCH); Secondary to drugs (drug-dependent or independent) | | Warm (mostly IgG, 37°C) | Primary: AIHA (idiopathic); Secondary to infection (Babesia); Secondary to drugs (e.g., cefotetan, ceftriaxone, piperacillin, methyldopa, fludarabine) |


6. INVESTIGATIONS & DIAGNOSIS

Diagnostic approach:

  1. Initial screening: CBC, reticulocyte count, bilirubin, LDH, haptoglobin.
  2. Site determination: Hemoglobinuria (intravascular) vs. unconjugated bilirubin (extravascular).
  3. Direct testing: Osmotic fragility (HS), EMA-binding test (membrane defects), G6PD activity assay.
  4. Advanced testing: Flow cytometry for PNH, molecular genetic analysis for inherited disorders.
  5. Imaging: Ultrasound for splenomegaly, liver iron quantification in chronic hemolysis.

6.1 Diagnostic Algorithms

Step-by-step diagnostic pathway:

• Step 1: Confirm hemolysis via reticulocyte count (>2% or >150,000/μL) and LDH/haptoglobin ratio. • Step 2: Differentiate intravascular/extravascular hemolysis (hemoglobinuria vs. unconjugated bilirubin). • Step 3: Screen for inherited causes (osmotic fragility, EMA test, G6PD assay). • Step 4: Confirm genetic mutations in suspected cases via molecular analysis.


7. MANAGEMENT & TREATMENT

Treatment strategies:

Hereditary spherocytosis: Splenectomy for severe cases (Hb <8 g/dL or symptomatic); folic acid supplementation. • Pyruvate kinase deficiency: Mitapivat (10 mg/kg/day), blood transfusions in severe cases (Hb <6 g/dL), folic acid (5 mg/day). • G6PD deficiency: Avoid oxidant triggers (fava beans, specific drugs like primaquine or dapsone). • Iron overload management: Phlebotomy or chelation therapy for secondary hemochromatosis. • Aplastic crisis: Blood transfusion for parvovirus B19-induced crises.

7.1 Treatment of Hereditary Spherocytosis

Management guidelines:

• Splenectomy: Indicated for hemoglobin <8 g/dL or symptomatic cases. • Post-splenectomy: Pneumococcal vaccination, lifelong penicillin prophylaxis. • Alternative therapies: hydroxyurea in refractory cases.

7.2 Treatment of Pyruvate Kinase Deficiency

Therapeutic options:

Mitapivat: 10 mg/kg/day (allosteric PK activator). • Blood transfusions: For severe anemia (Hb <6 g/dL). • Folic acid supplementation: 5 mg/day to support erythropoiesis.

7.3 Treatment of Paroxysmal Nocturnal Hemoglobinuria (PNH)

Management pathways based on complement inhibition:

C5 Blockade (e.g., Eculizumab): ◦ Mechanism: Prevents MAC formation. ◦ Outcome: Reduces intravascular hemolysis; however, some extravascular hemolysis may persist if blockade is incomplete. • C3 Blockade (e.g., Pegcetacoplan): ◦ Mechanism: Prevents C3 cleavage into C3b. ◦ Outcome: Prevents both MAC formation and opsonization by C3b; provides more robust protection against hemolysis.


8. PROGNOSIS & COMPLICATIONS

Long-term outcomes:

Inherited HAs: Variable prognosis based on severity and treatment response. • Complications: ◦ Iron overload (liver cirrhosis, heart failure) from chronic extravascular hemolysis. ◦ Thromboembolism: High risk in stomatocytosis; splenectomy is contraindicated. ◦ Aplastic crisis: Triggered by Parvovirus B19 in patients with chronic hemolysis (e.g., PK deficiency). • Mortality: Rare with modern management but significant in untreated severe cases.


9. SPECIAL CONSIDERATIONS

Key considerations:

Pregnancy: Increased hemolysis risk due to folate deficiency and placental demands. • Infections: Parvovirus B19, malaria, and viral hepatitis as triggers. • Drug interactions: Antimalarials (e.g., primaquine) in G6PD deficiency.

9.1 Pregnancy and Infection

Pregnancy-specific management:

• Monitor for decompensation (Hb <8 g/dL, reticulocyte count >5%). • Folic acid supplementation (4 mg/day) to prevent megaloblastic changes. • Avoid splenectomy in early pregnancy due to thromboembolism risk.


10. KEY PEARLS & CLINICAL TRAPS

Critical pearls:

Reticulocyte Count: Retic >2% or absolute >150,000/μL confirms hemolysis. • Osmotic Fragility: Diagnostic for HS but not specific for other membrane defects. • G6PD Safety: Must be ruled out before using oxidant drugs (e.g., primaquine, sulfonamides). • Stomatocytosis Warning: Splenectomy is strictly contraindicated due to high thromboembolic risk. • Intravascular Markers: Hemoglobinuria and low haptoglobin indicate intravascular hemolysis.


Reference Tables

TABLE 105-2 Features Common to Most Patients with a Hemolytic Disorder

Harrison's 22e, p.791

105 Hemolytic Anemias
Lucio Luzzatto, Lucia De Franceschi

TABLE 105-2 Features Common to Most Patients with a Hemolytic Disorder

GENERAL EXAMINATION JAUNDICE, PALLOR
Other physical findings Spleen may be enlarged; bossing of skull in severe
congenital cases
MCV, MCH Usually increased
Bilirubin Almost always increased (mostly unconjugated)
Haptoglobin Reduced to absent if hemolysis is at least in part
intravascular

TABLE 105-1 Classification of Hemolytic Anemias a Inherited

Harrison's 22e, p.791

INTRACORPUSCULAR DEFECTS EXTRACORPUSCULAR
FACTORS
Inherited Hemoglobinopathies
Enzymopathies
Membrane-cytoskeletal defects
Familial (atypical) hemolytic-
uremic syndrome
Paroxysmal nocturnal
hemoglobinuria (PNH)

TABLE 105-3 Inherited Diseases of the Red Cell Membrane-Cytoskeleton Complex

Harrison's 22e, p.794

GENE CHROMOSOMAL
LOCATION
PROTEIN PRODUCED DISEASE(S) WITH CERTAIN
MUTATIONS (INHERITANCE)
COMMENTS
SPTA1 1q22-q23 α-Spectrin HS (recessive) Rare
HE (dominant) Mutations of this gene account for about 65% of HE. More severe
forms may be due to coexistence of an otherwise silent mutant allele.
14q23-q24.1 β-Spectrin HS (dominant)
HE (dominant)
ANK1 8p11.2 Ankyrin HS (dominant) May account for majority of HS.
17q21 Band 3; also known as AE (anion
exchanger) or AE1
HS (dominant)
Southeast Asia ovalocytosis
(dominant)
Stomatocytosis
(cryohydrocytosis)
EPB41 1p33-p34.2 Band 4.1 HE (dominant) Mutations of this gene account for about 5% of HE, mostly with
prominent morphology but little/no hemolysis in heterozygotes; severe
hemolysis in homozygotes.
15q15-q21 Band 4.2 HS (recessive)
RHAG 6p21.1-p11 Rhesus-associated glycoprotein Chronic nonspherocytic
hemolytic anemia (recessive)
Very rare; associated with total loss of all Rh antigens.
One specific mutation in this gene entails loss of stomatin from the
cell membrane, causing overhydrated stomatocytosis.
16q23-q24 PIEZO1 (mechanosensitive ion
channel component 1 channel)
Dehydrated hereditary
stomatocytosis (dominant)
KCNN4 19q13.31 KCNN4
Intermediate conductance
calcium-activated potassium
channel protein 4 (Gardos
channel)
Dehydrated hereditary
stomatocytosis (dominant)
Clinical presentation similar to that of PIEZO1 mutants.
2q35-q36 ATP-binding cassette subfamily
B member 6
Familial pseudohyperkalemia
(dominant)
SLC2A1 1p34.2 GLUT1 glucose transporter Overhydrated hereditary
stomatocytosis
Associated with serious neurologic manifestations.
ANK1
EPB42
SPTB SPTA1
SLC2A1
RHAG
PIEZ01
KCNN4
ABCB6

TABLE 105-4 Red Cell Enzyme Abnormalities Causing Hemolysis

Harrison's 22e, p.796

ENZYME (ACRONYM) GENE SYMBOL;
CHROMOSOMAL
LOCATION
PREVALENCE OF ENZYME
DEFICIENCY (RANK)
CLINICAL
MANIFESTATIONS
EXTRA-RED CELL
COMMENTS
Glycolytic Pathway
Hexokinase (HK) HK1; 10q22 Very rare May benefit from splenectomy; BMTc
Glucose-6-phosphate isomerase (G6PI) GPI; 19q31.1 Rare (4); at least 60 cases
reporteda
NM, CNS May benefit from splenectomy
Phosphofructokinase (PFK)b PFKM; 12q13 Very rare Myopathy;
myoglobinuria
Aldolase ALDOA; 16q22-24 Very rare Myopathy
Triose phosphate isomerase (TPI) TPI1; 12p13.31 Very rare CNS (severe), NM
Glyceraldehyde 3-phosphate
dehydrogenase (GAPD)
GAPDH; 12p13.31 Very rare Myopathy
Bisphosphoglycerate mutase (BPGM) BPGM; 7q33 Very rare Erythrocytosis rather than hemolysis;
some of the rare mutations are in the
enzyme active site
Phosphoglycerate kinase (PGK) PGK1; Xq21.1 Very rare CNS, NM May benefit from splenectomy; BMTc
Pyruvate kinase (PK) PKLR; 1q22 Rare (2)a May benefit from splenectomy; BMTc
Redox
G6PD; Xq28
GSS; 20q11.22
GSR; 8p12
GCLC; 6p12.1
CYB5R3; 22q13.2
Common (1)a
Very rare
Very rare
Very rare
Rare
Very rarely granulocytes
CNS
Cataracts
CNS
CNS
Nucleotide Metabolism
Adenylate kinase (AK) AK1; 9q34.11 Very rare CNS May benefit from splenectomy
Pyrimidine 5’ nucleotidase (P5N) NTSC3A; 7p14.3 Rare (3)a May benefit from splenectomy

TABLE 105-5 Current World Health Organization Classification of Glucose 6-Phosphate Dehydrogenase (G6PD) Variants

Harrison's 22e, p.798

G6PD
VARIANT
CLASS
MEDIAN OF G6PD
ACTIVITY (% OF
NORMAL)
ASSOCIATED CLINICAL MANIFESTATIONS
Aa <20%b Chronic hemolytic anemia
<45%
Ca >60% None reported
Any

TABLE 105-6 Drugs That Carry Risk of Clinical Hemolysis in Persons with Glucose 6-Phosphate Dehydrogenase Deficiency

Harrison's 22e, p.799

DRUG CLASS
HIGH MEDIUM TO LOW
Antimalarials Primaquine
Tafenoquine
Chloroquine
Hydroxychloroquine
Quinine
Dapsone Sulfadimidine
Sulfamethoxazole
Sulfasalazine
Antibacterial/
antibiotics
Chloramphenicol
Ciprofloxacin
Cotrimoxazole
Nalidixic acid
Nitrofurantoin
Norfloxacin
p-Aminosalicylic acid
Niridazole
Antipyretic/
analgesics
Acetylsalicylic
acid high dose
(>3 g/d)
Acetaminophen
Acetanilide
Phenacetin
Phenazopyridine
Rasburicase
Pegloticase
Methylene blue
Toluidine blue
Ascorbic acid (>1 g)
Doxorubicin
Probenecid
Vitamin K analogues

TABLE 105-7 Diseases and Clinical Situations in Which Hemolysis Is Largely Intravascular Mismatched blood transfusion…

Harrison's 22e, p.801

ONSET/TIME COURSE MAIN MECHANISM APPROPRIATE
DIAGNOSTIC PROCEDURE
COMMENTS
Mismatched blood
transfusion
Abrupt Nearly always ABO
incompatibility
Repeat cross-match
Chronic with acute
exacerbations
Complement (C)-mediated
destruction of CD59(−) red cells
Flow cytometry to display a
CD59(−) red cell population
Paroxysmal cold
hemoglobinuria (PCH)
Acute Immune lysis of normal red
cells
Test for Donath-Landsteiner
antibody
Often triggered by viral infection
Very acute Exotoxins produced by
Clostridium perfringens
Blood cultures
Microangiopathic Acute or chronic Red cell fragmentation Red cell morphology on
blood smear
Different causes ranging from endothelial damage to
hemangioma to leaky prosthetic heart valve
Abrupt Mechanical destruction Targeted history taking
Favism Acute Destruction of older fraction of
G6PD-deficient red cells
G6PD assay Triggered by ingestion of large dish of fava beansa

TABLE 105-8 Classification of Acquired Immune Hemolytic Anemias

Harrison's 22e, p.802

CLINICAL SETTING TYPE OF ANTIBODY
COLD, MOSTLY
IgM, OPTIMAL
TEMPERATURE
4°C–30°C
WARM, MOSTLY IgG, OPTIMAL
TEMPERATURE 37°C; OR MIXED
Primary CAD AIHA (idiopathic)
EBV
CMV
Other
Secondary to other
infection
Mycoplasma
infection:
paroxysmal cold
hemoglobinuria
Babesia
CAD in:
Waldenström’s
disease
Lymphoma
Secondary to
drugs: drug-
induced immune
hemolytic anemia
Small minority (e.g.,
with lenalidomide)
Majority: currently most common
culprit drugs are cefotetan,
ceftriaxone, piperacillin,
methyldopa, fludarabine
Drug-dependent: antibody destroys red cells only when
drug present (e.g., rarely penicillin)
Drug-independent: antibody can destroy red cells even
when drug no longer present (e.g., methyldopa)