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Paroxysmal Nocturnal Haemoglobinuria (PNH)
Pathophysiology
X-linked PIG-A gene mutation —> deficiency of GPI-anchored proteins (CD55 & CD59)
Normal role of CD55 is to block action of C3 convertases on red cells
Lack of CD55/CD59 —> red cells vulnerable to MAC —> chronic haemolysis
Clinical Features
Intravascular Haemolysis
—> Large amounts of free Hb —> consumption of NO —> abdo pain, oesophageal spasm, erectile dysfunction and pulmonary hypertension
Thrombosis
Most common cause of death in PNH, occurs in 40% of patients, often unusual sites.
PNH is the highest risk thrombophilia
Panyctopenia
Associated with aplastic anaemia
Renal Failure
Renal vein thrombosis, ATN, siderosis of kidney
Who to Test (national PNH website)
Haemoglobinuria
DAT negative intravascular haemolysis
Unexplained haemolysis
Unexplained thrombosis
Either at unusual site – e.g. Budd-Chiari / intra-abdo, cerebral, dermal
Or in presence of haemolysis or cytopenias
MDS
Aplastic Anaemia
Other unexplained cytopenia
Diagnosis
Flow cytometry of peripheral blood for GPI-anchor deficiency
First test white cells to quantify the clone
2 Stages
Use markers to identify two WBC populations (not GPI-linked)
Neut – CD45 / CD15
Mono – CD45 / CD64
Then use 2 markers to identify deficient clone (GPI-linked)
Neut – FLAER / CD24 / CD66b
Mono – FLAER / CD14
Then if PNH clone found, test red cells to identify nature of deficiency (Type 2 vs 3)
2 Stages
Use CD235a (Glycophorin A) to identify red cell pop. (not GPI-linked)
Then use GPI-linked marker to find deficient pop. (CD59 preferred to CD55)
Results
Type 1 RBC – normal
Type 2 RBC – partial deficiency, may merge into the type 1 plots
Type 3 RBC – full deficiency
(N.B. WBC are used to identify the clone as false negatives occur with red cell tests due to short lifespan of the PNH cells, or recent transfusion of normal RBC’s.)
(N.B. FLAER technique – modified bacterial protein that binds GPI anchors – i.e. quantifies the non-PNH cells.)
CD5 Inhibitors (Eculizumab & ravulizumab)
Mechanism
Anti-C5 antibody, prevents C5 cleavage into C5a and C5b
Prevents MAC from causing intravascular red cell haemolysis
Prevents thrombosis & haemolysis, prolongs survival, prevents LT complications
However, C3 continues to be produced —> ongoing long-term extravascular haemolysis
Does not affect the underlying disease process of clonal haematopoeisis
Indications in UK (National PNH Service Website)
Thrombosis related to PNH
Complications of haemolysis (Renal impairment or Pul Hypertension)
Pregnancy (and up to 3 months post-partum)
Symptomatic Haemolysis (LDH >1.5x ULN with Hb <90)
Exceptional cases agreed with national commissioning services
Dose
Ravulizumab (NICE 2021) - IV infusion, weight-based, every 8 weeks
Eculizumab - IV infusion, 600mg weekly for first 5 weeks then 900mg fortnightly
Prior to use
Must give meningococcal vaccination prior to drug being released for use
Side Effects
Hyposplenic infection risks
If stopped suddenly —> severe drop in Hb. The PNH cells that had been surviving suddenly at risk of simultaneous breakdown.
AXLN1210 Trials 2019
301 Study - Rav vs Ecu in C5 inhibitor naïve patients. Demonstrated non-inferiority.
302 Study - Rav vs Ecu in C5 inhibitor experienced patients. Demonstrated non-inferiority.
Eculizumab, 2 weekly infusion
50% patients became transfusion independent (versus 0% of placebo group)
LDH improved
Emerging therapies
Iptacopan - oral complement factor B inhibitor. Achieved transfusion independence through resolution of extravascular haemolysis (Apply-PNH Phase III trial, ASH 2022)
Other ‘Complementopathies’
Atypical Haemolytic Uraemic Syndrome (aHUS)
Acts around alternative pathway
Caused by mutations/deficiencies of proteins that inhibit the alternative pathway
Results in constant firing of the complement cascade
Extremely difficult to differentiate from TTP.
33% of patients die or develop ESRF after 1st episode
Cold Agglutinin Disease (CHAD)
Classical pathway, starts with C1
Recognised as a clonal LPD
Unresponsive to steroids/splenectomy (as haemolysis is occurring in the liver)
Rx: Rituximab
Anti-C1 antibodies in development.